[{"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","citation":{"chicago":"Zhao, Ziyu, Irene Vercellino, Jana Knoppová, Roman Sobotka, James W. Murray, Peter J. Nixon, Leonid A Sazanov, and Josef Komenda. “The Ycf48 Accessory Factor Occupies the Site of the Oxygen-Evolving Manganese Cluster during Photosystem II Biogenesis.” <i>Nature Communications</i>. Springer Nature, 2023. <a href=\"https://doi.org/10.1038/s41467-023-40388-6\">https://doi.org/10.1038/s41467-023-40388-6</a>.","ista":"Zhao Z, Vercellino I, Knoppová J, Sobotka R, Murray JW, Nixon PJ, Sazanov LA, Komenda J. 2023. The Ycf48 accessory factor occupies the site of the oxygen-evolving manganese cluster during photosystem II biogenesis. Nature Communications. 14, 4681.","apa":"Zhao, Z., Vercellino, I., Knoppová, J., Sobotka, R., Murray, J. W., Nixon, P. J., … Komenda, J. (2023). The Ycf48 accessory factor occupies the site of the oxygen-evolving manganese cluster during photosystem II biogenesis. <i>Nature Communications</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41467-023-40388-6\">https://doi.org/10.1038/s41467-023-40388-6</a>","mla":"Zhao, Ziyu, et al. “The Ycf48 Accessory Factor Occupies the Site of the Oxygen-Evolving Manganese Cluster during Photosystem II Biogenesis.” <i>Nature Communications</i>, vol. 14, 4681, Springer Nature, 2023, doi:<a href=\"https://doi.org/10.1038/s41467-023-40388-6\">10.1038/s41467-023-40388-6</a>.","ama":"Zhao Z, Vercellino I, Knoppová J, et al. The Ycf48 accessory factor occupies the site of the oxygen-evolving manganese cluster during photosystem II biogenesis. <i>Nature Communications</i>. 2023;14. doi:<a href=\"https://doi.org/10.1038/s41467-023-40388-6\">10.1038/s41467-023-40388-6</a>","ieee":"Z. Zhao <i>et al.</i>, “The Ycf48 accessory factor occupies the site of the oxygen-evolving manganese cluster during photosystem II biogenesis,” <i>Nature Communications</i>, vol. 14. Springer Nature, 2023.","short":"Z. Zhao, I. Vercellino, J. Knoppová, R. Sobotka, J.W. Murray, P.J. Nixon, L.A. Sazanov, J. Komenda, Nature Communications 14 (2023)."},"language":[{"iso":"eng"}],"oa":1,"article_number":"4681","file":[{"file_id":"14044","date_updated":"2023-08-14T07:01:12Z","creator":"dernst","date_created":"2023-08-14T07:01:12Z","file_size":2315325,"checksum":"3b9043df3d51c300f9be95eac3ff9d0b","relation":"main_file","access_level":"open_access","content_type":"application/pdf","success":1,"file_name":"2023_NatureComm_Zhao.pdf"}],"department":[{"_id":"LeSa"}],"month":"08","file_date_updated":"2023-08-14T07:01:12Z","publication_status":"published","publication_identifier":{"eissn":["2041-1723"]},"acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"LifeSc"},{"_id":"ScienComp"}],"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"abstract":[{"lang":"eng","text":"Robust oxygenic photosynthesis requires a suite of accessory factors to ensure efficient assembly and repair of the oxygen-evolving photosystem two (PSII) complex. The highly conserved Ycf48 assembly factor binds to the newly synthesized D1 reaction center polypeptide and promotes the initial steps of PSII assembly, but its binding site is unclear. Here we use cryo-electron microscopy to determine the structure of a cyanobacterial PSII D1/D2 reaction center assembly complex with Ycf48 attached. Ycf48, a 7-bladed beta propeller, binds to the amino-acid residues of D1 that ultimately ligate the water-oxidising Mn4CaO5 cluster, thereby preventing the premature binding of Mn2+ and Ca2+ ions and protecting the site from damage. Interactions with D2 help explain how Ycf48 promotes assembly of the D1/D2 complex. Overall, our work provides valuable insights into the early stages of PSII assembly and the structural changes that create the binding site for the Mn4CaO5 cluster."}],"intvolume":"        14","has_accepted_license":"1","date_created":"2023-08-13T22:01:13Z","article_type":"original","volume":14,"oa_version":"Published Version","title":"The Ycf48 accessory factor occupies the site of the oxygen-evolving manganese cluster during photosystem II biogenesis","scopus_import":"1","day":"04","author":[{"last_name":"Zhao","full_name":"Zhao, Ziyu","first_name":"Ziyu"},{"id":"3ED6AF16-F248-11E8-B48F-1D18A9856A87","full_name":"Vercellino, Irene","last_name":"Vercellino","first_name":"Irene","orcid":"0000-0001-5618-3449"},{"full_name":"Knoppová, Jana","last_name":"Knoppová","first_name":"Jana"},{"last_name":"Sobotka","full_name":"Sobotka, Roman","first_name":"Roman"},{"first_name":"James W.","last_name":"Murray","full_name":"Murray, James W."},{"first_name":"Peter J.","last_name":"Nixon","full_name":"Nixon, Peter J."},{"orcid":"0000-0002-0977-7989","first_name":"Leonid A","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov"},{"full_name":"Komenda, Josef","last_name":"Komenda","first_name":"Josef"}],"date_published":"2023-08-04T00:00:00Z","acknowledgement":"P.J.N. and J.W.M. are grateful for the support of the Biotechnology & Biological Sciences Research Council (awards BB/L003260/1 and BB/P00931X/1). J. Knoppová, R.S. and J. Komenda were supported by the Czech Science Foundation (project 19-29225X) and by ERC project Photoredesign (no. 854126) and L.A.S. was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Electron Microscopy Facility (EMF), the Life Science Facility (LSF) and the IST high-performance computing cluster.","publication":"Nature Communications","status":"public","external_id":{"isi":["001042606700004"]},"isi":1,"year":"2023","quality_controlled":"1","ddc":["570"],"type":"journal_article","_id":"14040","date_updated":"2023-12-13T12:06:56Z","publisher":"Springer Nature","article_processing_charge":"Yes","doi":"10.1038/s41467-023-40388-6"},{"status":"public","publication":"The Biochemical Journal","pmid":1,"date_published":"2023-03-15T00:00:00Z","isi":1,"year":"2023","external_id":{"pmid":["36920092"],"isi":["000957065700001"]},"page":"319-333","ddc":["570"],"main_file_link":[{"url":"https://doi.org/10.1042/BCJ20210285","open_access":"1"}],"quality_controlled":"1","article_processing_charge":"No","doi":"10.1042/BCJ20210285","publisher":"Portland Press","_id":"12757","date_updated":"2023-08-01T13:45:12Z","type":"journal_article","oa":1,"language":[{"iso":"eng"}],"citation":{"ista":"Sazanov LA. 2023. From the ‘black box’ to ‘domino effect’ mechanism: What have we learned from the structures of respiratory complex I. The Biochemical Journal. 480(5), 319–333.","chicago":"Sazanov, Leonid A. “From the ‘black Box’ to ‘Domino Effect’ Mechanism: What Have We Learned from the Structures of Respiratory Complex I.” <i>The Biochemical Journal</i>. Portland Press, 2023. <a href=\"https://doi.org/10.1042/BCJ20210285\">https://doi.org/10.1042/BCJ20210285</a>.","mla":"Sazanov, Leonid A. “From the ‘black Box’ to ‘Domino Effect’ Mechanism: What Have We Learned from the Structures of Respiratory Complex I.” <i>The Biochemical Journal</i>, vol. 480, no. 5, Portland Press, 2023, pp. 319–33, doi:<a href=\"https://doi.org/10.1042/BCJ20210285\">10.1042/BCJ20210285</a>.","apa":"Sazanov, L. A. (2023). From the “black box” to “domino effect” mechanism: What have we learned from the structures of respiratory complex I. <i>The Biochemical Journal</i>. Portland Press. <a href=\"https://doi.org/10.1042/BCJ20210285\">https://doi.org/10.1042/BCJ20210285</a>","ama":"Sazanov LA. From the “black box” to “domino effect” mechanism: What have we learned from the structures of respiratory complex I. <i>The Biochemical Journal</i>. 2023;480(5):319-333. doi:<a href=\"https://doi.org/10.1042/BCJ20210285\">10.1042/BCJ20210285</a>","short":"L.A. Sazanov, The Biochemical Journal 480 (2023) 319–333.","ieee":"L. A. Sazanov, “From the ‘black box’ to ‘domino effect’ mechanism: What have we learned from the structures of respiratory complex I,” <i>The Biochemical Journal</i>, vol. 480, no. 5. Portland Press, pp. 319–333, 2023."},"issue":"5","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","month":"03","department":[{"_id":"LeSa"}],"has_accepted_license":"1","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"abstract":[{"lang":"eng","text":"My group and myself have studied respiratory complex I for almost 30 years, starting in 1994 when it was known as a L-shaped giant ‘black box' of bioenergetics. First breakthrough was the X-ray structure of the peripheral arm, followed by structures of the membrane arm and finally the entire complex from Thermus thermophilus. The developments in cryo-EM technology allowed us to solve the first complete structure of the twice larger, ∼1 MDa mammalian enzyme in 2016. However, the mechanism coupling, over large distances, the transfer of two electrons to pumping of four protons across the membrane remained an enigma. Recently we have solved high-resolution structures of mammalian and bacterial complex I under a range of redox conditions, including catalytic turnover. This allowed us to propose a robust and universal mechanism for complex I and related protein families. Redox reactions initially drive conformational changes around the quinone cavity and a long-distance transfer of substrate protons. These set up a stage for a series of electrostatically driven proton transfers along the membrane arm (‘domino effect'), eventually resulting in proton expulsion from the distal antiporter-like subunit. The mechanism radically differs from previous suggestions, however, it naturally explains all the unusual structural features of complex I. In this review I discuss the state of knowledge on complex I, including the current most controversial issues."}],"intvolume":"       480","publication_status":"published","publication_identifier":{"issn":["0264-6021"],"eissn":["1470-8728"]},"day":"15","scopus_import":"1","author":[{"first_name":"Leonid A","orcid":"0000-0002-0977-7989","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov"}],"oa_version":"Published Version","title":"From the 'black box' to 'domino effect' mechanism: What have we learned from the structures of respiratory complex I","volume":480,"date_created":"2023-03-26T22:01:06Z","article_type":"review"},{"file_date_updated":"2022-08-05T05:56:03Z","publication_status":"published","publication_identifier":{"issn":["0959-440X"]},"abstract":[{"text":"Complex I is one of the major respiratory complexes, conserved from bacteria to mammals. It oxidises NADH, reduces quinone and pumps protons across the membrane, thus playing a central role in the oxidative energy metabolism. In this review we discuss our current state of understanding the structure of complex I from various species of mammals, plants, fungi, and bacteria, as well as of several complex I-related proteins. By comparing the structural evidence from these systems in different redox states and data from mutagenesis and molecular simulations, we formulate the mechanisms of electron transfer and proton pumping and explain how they are conformationally and electrostatically coupled. Finally, we discuss the structural basis of the deactivation phenomenon in mammalian complex I.","lang":"eng"}],"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"intvolume":"        74","has_accepted_license":"1","date_created":"2022-04-15T09:32:35Z","article_type":"original","volume":74,"title":"Structure of respiratory complex I – An emerging blueprint for the mechanism","oa_version":"Published Version","scopus_import":"1","day":"01","author":[{"first_name":"Domen","last_name":"Kampjut","full_name":"Kampjut, Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Leonid A","orcid":"0000-0002-0977-7989","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov"}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"ama":"Kampjut D, Sazanov LA. Structure of respiratory complex I – An emerging blueprint for the mechanism. <i>Current Opinion in Structural Biology</i>. 2022;74. doi:<a href=\"https://doi.org/10.1016/j.sbi.2022.102350\">10.1016/j.sbi.2022.102350</a>","short":"D. Kampjut, L.A. Sazanov, Current Opinion in Structural Biology 74 (2022).","ieee":"D. Kampjut and L. A. Sazanov, “Structure of respiratory complex I – An emerging blueprint for the mechanism,” <i>Current Opinion in Structural Biology</i>, vol. 74. Elsevier, 2022.","ista":"Kampjut D, Sazanov LA. 2022. Structure of respiratory complex I – An emerging blueprint for the mechanism. Current Opinion in Structural Biology. 74, 102350.","chicago":"Kampjut, Domen, and Leonid A Sazanov. “Structure of Respiratory Complex I – An Emerging Blueprint for the Mechanism.” <i>Current Opinion in Structural Biology</i>. Elsevier, 2022. <a href=\"https://doi.org/10.1016/j.sbi.2022.102350\">https://doi.org/10.1016/j.sbi.2022.102350</a>.","mla":"Kampjut, Domen, and Leonid A. Sazanov. “Structure of Respiratory Complex I – An Emerging Blueprint for the Mechanism.” <i>Current Opinion in Structural Biology</i>, vol. 74, 102350, Elsevier, 2022, doi:<a href=\"https://doi.org/10.1016/j.sbi.2022.102350\">10.1016/j.sbi.2022.102350</a>.","apa":"Kampjut, D., &#38; Sazanov, L. A. (2022). Structure of respiratory complex I – An emerging blueprint for the mechanism. <i>Current Opinion in Structural Biology</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.sbi.2022.102350\">https://doi.org/10.1016/j.sbi.2022.102350</a>"},"language":[{"iso":"eng"}],"oa":1,"article_number":"102350","file":[{"checksum":"72bdde48853643a32d42b75f54965c44","relation":"main_file","content_type":"application/pdf","access_level":"open_access","file_name":"2022_CurrentOpStructBiology_Kampjut.pdf","success":1,"file_id":"11725","creator":"dernst","date_updated":"2022-08-05T05:56:03Z","date_created":"2022-08-05T05:56:03Z","file_size":815607}],"department":[{"_id":"LeSa"}],"month":"06","quality_controlled":"1","ddc":["570"],"type":"journal_article","_id":"11167","date_updated":"2023-08-03T06:31:06Z","publisher":"Elsevier","article_processing_charge":"Yes (via OA deal)","doi":"10.1016/j.sbi.2022.102350","date_published":"2022-06-01T00:00:00Z","pmid":1,"publication":"Current Opinion in Structural Biology","status":"public","keyword":["Molecular Biology","Structural Biology"],"external_id":{"isi":["000829029500020"],"pmid":["35316665"]},"year":"2022","isi":1},{"external_id":{"isi":["000815098500002"],"pmid":[" 35739187"]},"isi":1,"year":"2022","status":"public","publication":"Communications Biology","acknowledgement":"We thank Dr, Luke Formosa (Department of Biochemistry and Molecular Biology, Monash Biomedicine Discovery Institute, Monash University, Melbourne, Australia) for his valuable advice and assistance on NDUFA10 molecular studies and Dr. Francesc Canals and his team (Proteomics Laboratory, Vall d’Hebron Institute of Oncology [VHIO], Universitat Autònoma de Barcelona, Barcelona, Spain) for their assistance with LC-MS/MS analyses. This work was supported by the Spanish Ministry of Industry, Economy and Competitiveness [grants BFU2014-52618-R, SAF2017-87506, and PID2020-112929RB-I00 to Y.C.], by the Spanish Instituto de Salud Carlos III [grants PI21/00554 and PMP15/00025 to R.M.], co-financed by the European Regional Development Fund (ERDF), and by an NHMRC Project grant to M.R. (GNT1164459).\r\n","date_published":"2022-06-23T00:00:00Z","pmid":1,"publisher":"Springer Nature","doi":"10.1038/s42003-022-03568-6","article_processing_charge":"No","type":"journal_article","date_updated":"2023-08-03T11:51:58Z","_id":"11551","ddc":["570"],"quality_controlled":"1","month":"06","article_number":"620","file":[{"success":1,"file_name":"2022_communicationsbiology_Molina-Granada.pdf","access_level":"open_access","content_type":"application/pdf","relation":"main_file","checksum":"965f88bbcef3fd0c3e121340555c4467","file_size":2335369,"date_created":"2022-07-13T07:44:58Z","date_updated":"2022-07-13T07:44:58Z","creator":"kschuh","file_id":"11571"}],"department":[{"_id":"LeSa"}],"language":[{"iso":"eng"}],"oa":1,"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","issue":"1","citation":{"mla":"Molina-Granada, David, et al. “Most Mitochondrial DGTP Is Tightly Bound to Respiratory Complex I through the NDUFA10 Subunit.” <i>Communications Biology</i>, vol. 5, no. 1, 620, Springer Nature, 2022, doi:<a href=\"https://doi.org/10.1038/s42003-022-03568-6\">10.1038/s42003-022-03568-6</a>.","apa":"Molina-Granada, D., González-Vioque, E., Dibley, M. G., Cabrera-Pérez, R., Vallbona-Garcia, A., Torres-Torronteras, J., … Martí, R. (2022). Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. <i>Communications Biology</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s42003-022-03568-6\">https://doi.org/10.1038/s42003-022-03568-6</a>","ista":"Molina-Granada D, González-Vioque E, Dibley MG, Cabrera-Pérez R, Vallbona-Garcia A, Torres-Torronteras J, Sazanov LA, Ryan MT, Cámara Y, Martí R. 2022. Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. Communications Biology. 5(1), 620.","chicago":"Molina-Granada, David, Emiliano González-Vioque, Marris G. Dibley, Raquel Cabrera-Pérez, Antoni Vallbona-Garcia, Javier Torres-Torronteras, Leonid A Sazanov, Michael T. Ryan, Yolanda Cámara, and Ramon Martí. “Most Mitochondrial DGTP Is Tightly Bound to Respiratory Complex I through the NDUFA10 Subunit.” <i>Communications Biology</i>. Springer Nature, 2022. <a href=\"https://doi.org/10.1038/s42003-022-03568-6\">https://doi.org/10.1038/s42003-022-03568-6</a>.","short":"D. Molina-Granada, E. González-Vioque, M.G. Dibley, R. Cabrera-Pérez, A. Vallbona-Garcia, J. Torres-Torronteras, L.A. Sazanov, M.T. Ryan, Y. Cámara, R. Martí, Communications Biology 5 (2022).","ieee":"D. Molina-Granada <i>et al.</i>, “Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit,” <i>Communications Biology</i>, vol. 5, no. 1. Springer Nature, 2022.","ama":"Molina-Granada D, González-Vioque E, Dibley MG, et al. Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. <i>Communications Biology</i>. 2022;5(1). doi:<a href=\"https://doi.org/10.1038/s42003-022-03568-6\">10.1038/s42003-022-03568-6</a>"},"title":"Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit","oa_version":"Published Version","author":[{"first_name":"David","last_name":"Molina-Granada","full_name":"Molina-Granada, David"},{"first_name":"Emiliano","last_name":"González-Vioque","full_name":"González-Vioque, Emiliano"},{"first_name":"Marris G.","full_name":"Dibley, Marris G.","last_name":"Dibley"},{"first_name":"Raquel","last_name":"Cabrera-Pérez","full_name":"Cabrera-Pérez, Raquel"},{"full_name":"Vallbona-Garcia, Antoni","last_name":"Vallbona-Garcia","first_name":"Antoni"},{"first_name":"Javier","full_name":"Torres-Torronteras, Javier","last_name":"Torres-Torronteras"},{"last_name":"Sazanov","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-0977-7989","first_name":"Leonid A"},{"full_name":"Ryan, Michael T.","last_name":"Ryan","first_name":"Michael T."},{"last_name":"Cámara","full_name":"Cámara, Yolanda","first_name":"Yolanda"},{"full_name":"Martí, Ramon","last_name":"Martí","first_name":"Ramon"}],"scopus_import":"1","day":"23","date_created":"2022-07-10T22:01:52Z","volume":5,"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"intvolume":"         5","abstract":[{"lang":"eng","text":"Imbalanced mitochondrial dNTP pools are known players in the pathogenesis of multiple human diseases. Here we show that, even under physiological conditions, dGTP is largely overrepresented among other dNTPs in mitochondria of mouse tissues and human cultured cells. In addition, a vast majority of mitochondrial dGTP is tightly bound to NDUFA10, an accessory subunit of complex I of the mitochondrial respiratory chain. NDUFA10 shares a deoxyribonucleoside kinase (dNK) domain with deoxyribonucleoside kinases in the nucleotide salvage pathway, though no specific function beyond stabilizing the complex I holoenzyme has been described for this subunit. We mutated the dNK domain of NDUFA10 in human HEK-293T cells while preserving complex I assembly and activity. The NDUFA10E160A/R161A shows reduced dGTP binding capacity in vitro and leads to a 50% reduction in mitochondrial dGTP content, proving that most dGTP is directly bound to the dNK domain of NDUFA10. This interaction may represent a hitherto unknown mechanism regulating mitochondrial dNTP availability and linking oxidative metabolism to DNA maintenance."}],"has_accepted_license":"1","publication_status":"published","publication_identifier":{"eissn":["23993642"]},"file_date_updated":"2022-07-13T07:44:58Z"},{"volume":23,"date_created":"2021-10-24T22:01:35Z","article_type":"original","scopus_import":"1","day":"01","author":[{"orcid":" 0000-0001-5618-3449","first_name":"Irene","id":"3ED6AF16-F248-11E8-B48F-1D18A9856A87","full_name":"Vercellino, Irene","last_name":"Vercellino"},{"orcid":"0000-0002-0977-7989","first_name":"Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","last_name":"Sazanov"}],"title":"The assembly, regulation and function of the mitochondrial respiratory chain","oa_version":"None","publication_identifier":{"eissn":["1471-0080"],"issn":["1471-0072"]},"publication_status":"published","abstract":[{"lang":"eng","text":"The mitochondrial oxidative phosphorylation system is central to cellular metabolism. It comprises five enzymatic complexes and two mobile electron carriers that work in a mitochondrial respiratory chain. By coupling the oxidation of reducing equivalents coming into mitochondria to the generation and subsequent dissipation of a proton gradient across the inner mitochondrial membrane, this electron transport chain drives the production of ATP, which is then used as a primary energy carrier in virtually all cellular processes. Minimal perturbations of the respiratory chain activity are linked to diseases; therefore, it is necessary to understand how these complexes are assembled and regulated and how they function. In this Review, we outline the latest assembly models for each individual complex, and we also highlight the recent discoveries indicating that the formation of larger assemblies, known as respiratory supercomplexes, originates from the association of the intermediates of individual complexes. We then discuss how recent cryo-electron microscopy structures have been key to answering open questions on the function of the electron transport chain in mitochondrial respiration and how supercomplexes and other factors, including metabolites, can regulate the activity of the single complexes. When relevant, we discuss how these mechanisms contribute to physiology and outline their deregulation in human diseases."}],"intvolume":"        23","department":[{"_id":"LeSa"}],"month":"02","citation":{"ieee":"I. Vercellino and L. A. Sazanov, “The assembly, regulation and function of the mitochondrial respiratory chain,” <i>Nature Reviews Molecular Cell Biology</i>, vol. 23. Springer Nature, pp. 141–161, 2022.","short":"I. Vercellino, L.A. Sazanov, Nature Reviews Molecular Cell Biology 23 (2022) 141–161.","ama":"Vercellino I, Sazanov LA. The assembly, regulation and function of the mitochondrial respiratory chain. <i>Nature Reviews Molecular Cell Biology</i>. 2022;23:141–161. doi:<a href=\"https://doi.org/10.1038/s41580-021-00415-0\">10.1038/s41580-021-00415-0</a>","apa":"Vercellino, I., &#38; Sazanov, L. A. (2022). The assembly, regulation and function of the mitochondrial respiratory chain. <i>Nature Reviews Molecular Cell Biology</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41580-021-00415-0\">https://doi.org/10.1038/s41580-021-00415-0</a>","mla":"Vercellino, Irene, and Leonid A. Sazanov. “The Assembly, Regulation and Function of the Mitochondrial Respiratory Chain.” <i>Nature Reviews Molecular Cell Biology</i>, vol. 23, Springer Nature, 2022, pp. 141–161, doi:<a href=\"https://doi.org/10.1038/s41580-021-00415-0\">10.1038/s41580-021-00415-0</a>.","chicago":"Vercellino, Irene, and Leonid A Sazanov. “The Assembly, Regulation and Function of the Mitochondrial Respiratory Chain.” <i>Nature Reviews Molecular Cell Biology</i>. Springer Nature, 2022. <a href=\"https://doi.org/10.1038/s41580-021-00415-0\">https://doi.org/10.1038/s41580-021-00415-0</a>.","ista":"Vercellino I, Sazanov LA. 2022. The assembly, regulation and function of the mitochondrial respiratory chain. Nature Reviews Molecular Cell Biology. 23, 141–161."},"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","language":[{"iso":"eng"}],"_id":"10182","date_updated":"2023-08-02T06:55:42Z","type":"journal_article","article_processing_charge":"No","doi":"10.1038/s41580-021-00415-0","publisher":"Springer Nature","quality_controlled":"1","page":"141–161","isi":1,"year":"2022","external_id":{"pmid":["34621061"],"isi":["000705697100001"]},"pmid":1,"date_published":"2022-02-01T00:00:00Z","status":"public","publication":"Nature Reviews Molecular Cell Biology"},{"file":[{"content_type":"application/pdf","access_level":"open_access","file_name":"EcCxI_manuscript_rev3_noSI_updated_withFigs_opt.pdf","success":1,"checksum":"d42a93e24f59e883ef0b5429832391d0","relation":"main_file","creator":"lsazanov","date_updated":"2023-05-30T17:05:31Z","date_created":"2023-05-30T17:05:31Z","file_size":1425655,"file_id":"13104"},{"file_id":"13105","date_created":"2023-05-30T17:07:05Z","file_size":9842513,"creator":"lsazanov","date_updated":"2023-05-30T17:07:05Z","relation":"main_file","checksum":"5422bc0a73b3daadafa262c7ea6deae3","file_name":"EcCxI_manuscript_rev3_SI_All_opt_upd.pdf","success":1,"content_type":"application/pdf","access_level":"open_access"}],"department":[{"_id":"LeSa"}],"month":"09","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"ieee":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, and L. A. Sazanov, “A universal coupling mechanism of respiratory complex I,” <i>Nature</i>, vol. 609, no. 7928. Springer Nature, pp. 808–814, 2022.","short":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, L.A. Sazanov, Nature 609 (2022) 808–814.","ama":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. A universal coupling mechanism of respiratory complex I. <i>Nature</i>. 2022;609(7928):808-814. doi:<a href=\"https://doi.org/10.1038/s41586-022-05199-7\">10.1038/s41586-022-05199-7</a>","apa":"Kravchuk, V., Petrova, O., Kampjut, D., Wojciechowska-Bason, A., Breese, Z., &#38; Sazanov, L. A. (2022). A universal coupling mechanism of respiratory complex I. <i>Nature</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41586-022-05199-7\">https://doi.org/10.1038/s41586-022-05199-7</a>","mla":"Kravchuk, Vladyslav, et al. “A Universal Coupling Mechanism of Respiratory Complex I.” <i>Nature</i>, vol. 609, no. 7928, Springer Nature, 2022, pp. 808–14, doi:<a href=\"https://doi.org/10.1038/s41586-022-05199-7\">10.1038/s41586-022-05199-7</a>.","chicago":"Kravchuk, Vladyslav, Olga Petrova, Domen Kampjut, Anna Wojciechowska-Bason, Zara Breese, and Leonid A Sazanov. “A Universal Coupling Mechanism of Respiratory Complex I.” <i>Nature</i>. Springer Nature, 2022. <a href=\"https://doi.org/10.1038/s41586-022-05199-7\">https://doi.org/10.1038/s41586-022-05199-7</a>.","ista":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. 2022. A universal coupling mechanism of respiratory complex I. Nature. 609(7928), 808–814."},"issue":"7928","language":[{"iso":"eng"}],"oa":1,"date_created":"2023-01-12T12:04:33Z","article_type":"original","volume":609,"title":"A universal coupling mechanism of respiratory complex I","oa_version":"Submitted Version","scopus_import":"1","day":"22","author":[{"first_name":"Vladyslav","last_name":"Kravchuk","id":"4D62F2A6-F248-11E8-B48F-1D18A9856A87","full_name":"Kravchuk, Vladyslav"},{"first_name":"Olga","last_name":"Petrova","id":"5D8C9660-5D49-11EA-8188-567B3DDC885E","full_name":"Petrova, Olga"},{"first_name":"Domen","full_name":"Kampjut, Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87","last_name":"Kampjut"},{"full_name":"Wojciechowska-Bason, Anna","last_name":"Wojciechowska-Bason","first_name":"Anna"},{"full_name":"Breese, Zara","last_name":"Breese","first_name":"Zara"},{"last_name":"Sazanov","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","first_name":"Leonid A","orcid":"0000-0002-0977-7989"}],"file_date_updated":"2023-05-30T17:07:05Z","publication_status":"published","publication_identifier":{"issn":["0028-0836"],"eissn":["1476-4687"]},"acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"LifeSc"},{"_id":"ScienComp"}],"abstract":[{"text":"Complex I is the first enzyme in the respiratory chain, which is responsible for energy production in mitochondria and bacteria1. Complex I couples the transfer of two electrons from NADH to quinone and the translocation of four protons across the membrane2, but the coupling mechanism remains contentious. Here we present cryo-electron microscopy structures of Escherichia coli complex I (EcCI) in different redox states, including catalytic turnover. EcCI exists mostly in the open state, in which the quinone cavity is exposed to the cytosol, allowing access for water molecules, which enable quinone movements. Unlike the mammalian paralogues3, EcCI can convert to the closed state only during turnover, showing that closed and open states are genuine turnover intermediates. The open-to-closed transition results in the tightly engulfed quinone cavity being connected to the central axis of the membrane arm, a source of substrate protons. Consistently, the proportion of the closed state increases with increasing pH. We propose a detailed but straightforward and robust mechanism comprising a ‘domino effect’ series of proton transfers and electrostatic interactions: the forward wave (‘dominoes stacking’) primes the pump, and the reverse wave (‘dominoes falling’) results in the ejection of all pumped protons from the distal subunit NuoL. This mechanism explains why protons exit exclusively from the NuoL subunit and is supported by our mutagenesis data. We contend that this is a universal coupling mechanism of complex I and related enzymes.","lang":"eng"}],"intvolume":"       609","has_accepted_license":"1","keyword":["Multidisciplinary"],"related_material":{"record":[{"relation":"dissertation_contains","status":"public","id":"12781"}],"link":[{"url":"https://doi.org/10.1038/s41586-022-05457-8","relation":"erratum"},{"url":"https://ista.ac.at/en/news/proton-dominos-kick-off-life/","description":"News on ISTA website","relation":"press_release"}]},"external_id":{"isi":["000854788200001"],"pmid":["36104567"]},"isi":1,"year":"2022","date_published":"2022-09-22T00:00:00Z","acknowledgement":"This research was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Electron Microscopy Facility (EMF), the Life Science Facility (LSF) and the IST high-performance computing cluster. We thank V.-V. Hodirnau from IST Austria EMF, M. Babiak from CEITEC for assistance with collecting cryo-EM data and A. Charnagalov for the assistance with protein purification. V.K. was a recipient of a DOC Fellowship of the Austrian Academy of Sciences at the Institute of Science and Technology, Austria. V.K. and O.P. are funded by the ERC Advanced Grant 101020697 RESPICHAIN to L.S. This work was also supported by the Medical Research Council (UK).","ec_funded":1,"pmid":1,"status":"public","publication":"Nature","project":[{"name":"Structural characterization of E. coli complex I: an important mechanistic model","grant_number":"25541","_id":"238A0A5A-32DE-11EA-91FC-C7463DDC885E"},{"_id":"627abdeb-2b32-11ec-9570-ec31a97243d3","grant_number":"101020697","name":"Structure and mechanism of respiratory chain molecular machines","call_identifier":"H2020"}],"type":"journal_article","_id":"12138","date_updated":"2023-08-04T08:54:52Z","publisher":"Springer Nature","article_processing_charge":"No","doi":"10.1038/s41586-022-05199-7","quality_controlled":"1","ddc":["572"],"page":"808-814"},{"article_processing_charge":"No","doi":"10.1073/pnas.2020857118","publisher":"National Academy of Sciences","_id":"8993","date_updated":"2023-08-07T13:29:23Z","type":"journal_article","main_file_link":[{"url":"https://doi.org/10.1073/pnas.2020857118","open_access":"1"}],"quality_controlled":"1","year":"2021","isi":1,"related_material":{"link":[{"relation":"erratum","url":"https://doi.org/10.1073/pnas.2102232118"}]},"external_id":{"isi":["000607270100073"],"pmid":["33443187"]},"publication":"PNAS","status":"public","project":[{"_id":"261099A6-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","grant_number":"742985","name":"Tracing Evolution of Auxin Transport and Polarity in Plants"}],"ec_funded":1,"pmid":1,"acknowledgement":"This work was supported by Austrian Science Fund Grant FWF P21533-B20 (to L.A.); German Research Foundation Grant DFG HA3468/6-1 (to U.Z.H.); and European Research Council Grant 742985 (to J.F.). We thank Herta Steinkellner and Alexandra Castilho for N. benthamiana plants, Fabian Nagelreiter for statistical advice, Lanassa Bassukas for help with [ɣ32P]-\r\nATP assays, and Josef Penninger for providing access to mass spectrometry instruments at the Vienna BioCenter Core Facilities. We thank PNAS reviewers for the many comments and suggestions that helped to improve this manuscript.","date_published":"2021-01-05T00:00:00Z","scopus_import":"1","day":"05","author":[{"first_name":"Lindy","last_name":"Abas","full_name":"Abas, Lindy"},{"first_name":"Martina","last_name":"Kolb","full_name":"Kolb, Martina"},{"full_name":"Stadlmann, Johannes","last_name":"Stadlmann","first_name":"Johannes"},{"full_name":"Janacek, Dorina P.","last_name":"Janacek","first_name":"Dorina P."},{"first_name":"Kristina","orcid":"0000-0003-1581-881X","full_name":"Lukic, Kristina","id":"2B04DB84-F248-11E8-B48F-1D18A9856A87","last_name":"Lukic"},{"first_name":"Claus","last_name":"Schwechheimer","full_name":"Schwechheimer, Claus"},{"last_name":"Sazanov","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","orcid":"0000-0002-0977-7989","first_name":"Leonid A"},{"first_name":"Lukas","full_name":"Mach, Lukas","last_name":"Mach"},{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","full_name":"Friml, Jiří","last_name":"Friml","orcid":"0000-0002-8302-7596","first_name":"Jiří"},{"last_name":"Hammes","full_name":"Hammes, Ulrich Z.","first_name":"Ulrich Z."}],"oa_version":"Published Version","title":"Naphthylphthalamic acid associates with and inhibits PIN auxin transporters","volume":118,"date_created":"2021-01-03T23:01:23Z","article_type":"original","intvolume":"       118","abstract":[{"text":"N-1-naphthylphthalamic acid (NPA) is a key inhibitor of directional (polar) transport of the hormone auxin in plants. For decades, it has been a pivotal tool in elucidating the unique polar auxin transport-based processes underlying plant growth and development. Its exact mode of action has long been sought after and is still being debated, with prevailing mechanistic schemes describing only indirect connections between NPA and the main transporters responsible for directional transport, namely PIN auxin exporters. Here we present data supporting a model in which NPA associates with PINs in a more direct manner than hitherto postulated. We show that NPA inhibits PIN activity in a heterologous oocyte system and that expression of NPA-sensitive PINs in plant, yeast, and oocyte membranes leads to specific saturable NPA binding. We thus propose that PINs are a bona fide NPA target. This offers a straightforward molecular basis for NPA inhibition of PIN-dependent auxin transport and a logical parsimonious explanation for the known physiological effects of NPA on plant growth, as well as an alternative hypothesis to interpret past and future results. We also introduce PIN dimerization and describe an effect of NPA on this, suggesting that NPA binding could be exploited to gain insights into structural aspects of PINs related to their transport mechanism.","lang":"eng"}],"publication_identifier":{"eissn":["10916490"],"issn":["00278424"]},"publication_status":"published","month":"01","department":[{"_id":"JiFr"},{"_id":"LeSa"}],"article_number":"e2020857118","oa":1,"language":[{"iso":"eng"}],"citation":{"ista":"Abas L, Kolb M, Stadlmann J, Janacek DP, Lukic K, Schwechheimer C, Sazanov LA, Mach L, Friml J, Hammes UZ. 2021. Naphthylphthalamic acid associates with and inhibits PIN auxin transporters. PNAS. 118(1), e2020857118.","chicago":"Abas, Lindy, Martina Kolb, Johannes Stadlmann, Dorina P. Janacek, Kristina Lukic, Claus Schwechheimer, Leonid A Sazanov, Lukas Mach, Jiří Friml, and Ulrich Z. Hammes. “Naphthylphthalamic Acid Associates with and Inhibits PIN Auxin Transporters.” <i>PNAS</i>. National Academy of Sciences, 2021. <a href=\"https://doi.org/10.1073/pnas.2020857118\">https://doi.org/10.1073/pnas.2020857118</a>.","apa":"Abas, L., Kolb, M., Stadlmann, J., Janacek, D. P., Lukic, K., Schwechheimer, C., … Hammes, U. Z. (2021). Naphthylphthalamic acid associates with and inhibits PIN auxin transporters. <i>PNAS</i>. National Academy of Sciences. <a href=\"https://doi.org/10.1073/pnas.2020857118\">https://doi.org/10.1073/pnas.2020857118</a>","mla":"Abas, Lindy, et al. “Naphthylphthalamic Acid Associates with and Inhibits PIN Auxin Transporters.” <i>PNAS</i>, vol. 118, no. 1, e2020857118, National Academy of Sciences, 2021, doi:<a href=\"https://doi.org/10.1073/pnas.2020857118\">10.1073/pnas.2020857118</a>.","ama":"Abas L, Kolb M, Stadlmann J, et al. Naphthylphthalamic acid associates with and inhibits PIN auxin transporters. <i>PNAS</i>. 2021;118(1). doi:<a href=\"https://doi.org/10.1073/pnas.2020857118\">10.1073/pnas.2020857118</a>","ieee":"L. Abas <i>et al.</i>, “Naphthylphthalamic acid associates with and inhibits PIN auxin transporters,” <i>PNAS</i>, vol. 118, no. 1. National Academy of Sciences, 2021.","short":"L. Abas, M. Kolb, J. Stadlmann, D.P. Janacek, K. Lukic, C. Schwechheimer, L.A. Sazanov, L. Mach, J. Friml, U.Z. Hammes, PNAS 118 (2021)."},"issue":"1","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8"},{"isi":1,"year":"2021","external_id":{"pmid":["33665558"],"isi":["000631646000012"]},"ec_funded":1,"pmid":1,"acknowledgement":"We thank the Electron Microscopy Facilities at the Institute of Science and Technology Austria and at the Vienna Biocenter for providing access and training for the electron microscopes. This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie Grant Agreement no. 665385 .","date_published":"2021-03-19T00:00:00Z","status":"public","publication":"iScience","project":[{"_id":"2564DBCA-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","name":"International IST Doctoral Program","grant_number":"665385"}],"_id":"9205","date_updated":"2023-08-07T13:54:06Z","type":"journal_article","article_processing_charge":"No","doi":"10.1016/j.isci.2021.102139","publisher":"Elsevier","quality_controlled":"1","ddc":["570"],"department":[{"_id":"LeSa"}],"file":[{"file_name":"2021_iScience_Kampjut.pdf","success":1,"content_type":"application/pdf","access_level":"open_access","relation":"main_file","checksum":"50585447386fe5842f07ab9b3a66e7e9","file_size":7431411,"date_created":"2021-03-03T07:38:14Z","creator":"dernst","date_updated":"2021-03-03T07:38:14Z","file_id":"9219"}],"article_number":"102139","month":"03","citation":{"ama":"Kampjut D, Steiner J, Sazanov LA. Cryo-EM grid optimization for membrane proteins. <i>iScience</i>. 2021;24(3). doi:<a href=\"https://doi.org/10.1016/j.isci.2021.102139\">10.1016/j.isci.2021.102139</a>","short":"D. Kampjut, J. Steiner, L.A. Sazanov, IScience 24 (2021).","ieee":"D. Kampjut, J. Steiner, and L. A. Sazanov, “Cryo-EM grid optimization for membrane proteins,” <i>iScience</i>, vol. 24, no. 3. Elsevier, 2021.","chicago":"Kampjut, Domen, Julia Steiner, and Leonid A Sazanov. “Cryo-EM Grid Optimization for Membrane Proteins.” <i>IScience</i>. Elsevier, 2021. <a href=\"https://doi.org/10.1016/j.isci.2021.102139\">https://doi.org/10.1016/j.isci.2021.102139</a>.","ista":"Kampjut D, Steiner J, Sazanov LA. 2021. Cryo-EM grid optimization for membrane proteins. iScience. 24(3), 102139.","mla":"Kampjut, Domen, et al. “Cryo-EM Grid Optimization for Membrane Proteins.” <i>IScience</i>, vol. 24, no. 3, 102139, Elsevier, 2021, doi:<a href=\"https://doi.org/10.1016/j.isci.2021.102139\">10.1016/j.isci.2021.102139</a>.","apa":"Kampjut, D., Steiner, J., &#38; Sazanov, L. A. (2021). Cryo-EM grid optimization for membrane proteins. <i>IScience</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.isci.2021.102139\">https://doi.org/10.1016/j.isci.2021.102139</a>"},"issue":"3","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","oa":1,"language":[{"iso":"eng"}],"volume":24,"date_created":"2021-02-28T23:01:24Z","article_type":"original","day":"19","scopus_import":"1","author":[{"first_name":"Domen","last_name":"Kampjut","id":"37233050-F248-11E8-B48F-1D18A9856A87","full_name":"Kampjut, Domen"},{"full_name":"Steiner, Julia","id":"3BB67EB0-F248-11E8-B48F-1D18A9856A87","last_name":"Steiner","first_name":"Julia"},{"first_name":"Leonid A","orcid":"0000-0002-0977-7989","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","last_name":"Sazanov"}],"title":"Cryo-EM grid optimization for membrane proteins","oa_version":"Published Version","file_date_updated":"2021-03-03T07:38:14Z","publication_identifier":{"eissn":["25890042"]},"publication_status":"published","has_accepted_license":"1","acknowledged_ssus":[{"_id":"EM-Fac"}],"abstract":[{"text":"Cryo-EM grid preparation is an important bottleneck in protein structure determination, especially for membrane proteins, typically requiring screening of a large number of conditions. We systematically investigated the effects of buffer components, blotting conditions and grid types on the outcome of grid preparation of five different membrane protein samples. Aggregation was the most common type of problem which was addressed by changing detergents, salt concentration or reconstitution of proteins into nanodiscs or amphipols. We show that the optimal concentration of detergent is between 0.05 and 0.4% and that the presence of a low concentration of detergent with a high critical micellar concentration protects the proteins from denaturation at the air-water interface. Furthermore, we discuss the strategies for achieving an adequate ice thickness, particle coverage and orientation distribution on free ice and on support films. Our findings provide a clear roadmap for comprehensive screening of conditions for cryo-EM grid preparation of membrane proteins.","lang":"eng"}],"license":"https://creativecommons.org/licenses/by-nc-nd/4.0/","tmp":{"image":"/images/cc_by_nc_nd.png","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode","short":"CC BY-NC-ND (4.0)"},"intvolume":"        24"},{"page":"364-367","quality_controlled":"1","doi":"10.1038/s41586-021-03927-z","article_processing_charge":"No","publisher":"Springer Nature","date_updated":"2023-08-14T08:01:21Z","_id":"10146","type":"journal_article","project":[{"grant_number":"754411","name":"ISTplus - Postdoctoral Fellowships","call_identifier":"H2020","_id":"260C2330-B435-11E9-9278-68D0E5697425"}],"publication":"Nature","status":"public","pmid":1,"ec_funded":1,"date_published":"2021-10-14T00:00:00Z","acknowledgement":"We thank the pre-clinical facility of the IST Austria and A. Venturino for assistance with the animals; and V.-V. Hodirnau for assistance during the Titan Krios data collection, performed at the IST Austria. The data processing was performed at the IST high-performance computing cluster. This project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement no. 754411.","year":"2021","isi":1,"external_id":{"isi":["000704581600001"],"pmid":["34616041"]},"related_material":{"link":[{"relation":"press_release","description":"News on IST Webpage","url":"https://ist.ac.at/en/news/boosting-the-cells-power-house/"}]},"intvolume":"       598","abstract":[{"lang":"eng","text":"The enzymes of the mitochondrial electron transport chain are key players of cell metabolism. Despite being active when isolated, in vivo they associate into supercomplexes1, whose precise role is debated. Supercomplexes CIII2CIV1-2 (refs. 2,3), CICIII2 (ref. 4) and CICIII2CIV (respirasome)5,6,7,8,9,10 exist in mammals, but in contrast to CICIII2 and the respirasome, to date the only known eukaryotic structures of CIII2CIV1-2 come from Saccharomyces cerevisiae11,12 and plants13, which have different organization. Here we present the first, to our knowledge, structures of mammalian (mouse and ovine) CIII2CIV and its assembly intermediates, in different conformations. We describe the assembly of CIII2CIV from the CIII2 precursor to the final CIII2CIV conformation, driven by the insertion of the N terminus of the assembly factor SCAF1 (ref. 14) deep into CIII2, while its C terminus is integrated into CIV. Our structures (which include CICIII2 and the respirasome) also confirm that SCAF1 is exclusively required for the assembly of CIII2CIV and has no role in the assembly of the respirasome. We show that CIII2 is asymmetric due to the presence of only one copy of subunit 9, which straddles both monomers and prevents the attachment of a second copy of SCAF1 to CIII2, explaining the presence of one copy of CIV in CIII2CIV in mammals. Finally, we show that CIII2 and CIV gain catalytic advantage when assembled into the supercomplex and propose a role for CIII2CIV in fine tuning the efficiency of electron transfer in the electron transport chain."}],"acknowledged_ssus":[{"_id":"PreCl"},{"_id":"EM-Fac"},{"_id":"ScienComp"}],"publication_identifier":{"eissn":["1476-4687"],"issn":["0028-0836"]},"publication_status":"published","author":[{"full_name":"Vercellino, Irene","id":"3ED6AF16-F248-11E8-B48F-1D18A9856A87","last_name":"Vercellino","orcid":"0000-0001-5618-3449","first_name":"Irene"},{"id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989"}],"day":"14","scopus_import":"1","title":"Structure and assembly of the mammalian mitochondrial supercomplex CIII<sub>2</sub>CIV","oa_version":"None","volume":598,"article_type":"original","date_created":"2021-10-17T22:01:17Z","language":[{"iso":"eng"}],"issue":"7880","citation":{"ama":"Vercellino I, Sazanov LA. Structure and assembly of the mammalian mitochondrial supercomplex CIII<sub>2</sub>CIV. <i>Nature</i>. 2021;598(7880):364-367. doi:<a href=\"https://doi.org/10.1038/s41586-021-03927-z\">10.1038/s41586-021-03927-z</a>","short":"I. Vercellino, L.A. Sazanov, Nature 598 (2021) 364–367.","ieee":"I. Vercellino and L. A. Sazanov, “Structure and assembly of the mammalian mitochondrial supercomplex CIII<sub>2</sub>CIV,” <i>Nature</i>, vol. 598, no. 7880. Springer Nature, pp. 364–367, 2021.","chicago":"Vercellino, Irene, and Leonid A Sazanov. “Structure and Assembly of the Mammalian Mitochondrial Supercomplex CIII<sub>2</sub>CIV.” <i>Nature</i>. Springer Nature, 2021. <a href=\"https://doi.org/10.1038/s41586-021-03927-z\">https://doi.org/10.1038/s41586-021-03927-z</a>.","ista":"Vercellino I, Sazanov LA. 2021. Structure and assembly of the mammalian mitochondrial supercomplex CIII<sub>2</sub>CIV. Nature. 598(7880), 364–367.","mla":"Vercellino, Irene, and Leonid A. Sazanov. “Structure and Assembly of the Mammalian Mitochondrial Supercomplex CIII<sub>2</sub>CIV.” <i>Nature</i>, vol. 598, no. 7880, Springer Nature, 2021, pp. 364–67, doi:<a href=\"https://doi.org/10.1038/s41586-021-03927-z\">10.1038/s41586-021-03927-z</a>.","apa":"Vercellino, I., &#38; Sazanov, L. A. (2021). Structure and assembly of the mammalian mitochondrial supercomplex CIII<sub>2</sub>CIV. <i>Nature</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41586-021-03927-z\">https://doi.org/10.1038/s41586-021-03927-z</a>"},"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","month":"10","department":[{"_id":"LeSa"}]},{"external_id":{"pmid":["32335026"],"isi":["000540842000012"]},"year":"2020","isi":1,"date_published":"2020-08-01T00:00:00Z","pmid":1,"publication":"Biochimica et Biophysica Acta - Bioenergetics","status":"public","type":"journal_article","date_updated":"2023-08-21T06:19:18Z","_id":"7788","publisher":"Elsevier","doi":"10.1016/j.bbabio.2020.148213","article_processing_charge":"No","quality_controlled":"1","ddc":["570"],"article_number":"148213","file":[{"creator":"dernst","date_updated":"2020-07-14T12:48:03Z","date_created":"2020-05-04T12:25:19Z","file_size":3826792,"file_id":"7798","access_level":"open_access","content_type":"application/pdf","file_name":"2020_BBA_Adjobo_Hermans.pdf","checksum":"a9b152381307cf45fe266a8dc5640388","relation":"main_file"}],"department":[{"_id":"LeSa"}],"month":"08","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","issue":"8","citation":{"ista":"Adjobo-Hermans MJW, De Haas R, Willems PHGM, Wojtala A, Van Emst-De Vries SE, Wagenaars JA, Van Den Brand M, Rodenburg RJ, Smeitink JAM, Nijtmans LG, Sazanov LA, Wieckowski MR, Koopman WJH. 2020. NDUFS4 deletion triggers loss of NDUFA12 in Ndufs4−/− mice and Leigh syndrome patients: A stabilizing role for NDUFAF2. Biochimica et Biophysica Acta - Bioenergetics. 1861(8), 148213.","chicago":"Adjobo-Hermans, Merel J.W., Ria De Haas, Peter H.G.M. Willems, Aleksandra Wojtala, Sjenet E. Van Emst-De Vries, Jori A. Wagenaars, Mariel Van Den Brand, et al. “NDUFS4 Deletion Triggers Loss of NDUFA12 in Ndufs4−/− Mice and Leigh Syndrome Patients: A Stabilizing Role for NDUFAF2.” <i>Biochimica et Biophysica Acta - Bioenergetics</i>. Elsevier, 2020. <a href=\"https://doi.org/10.1016/j.bbabio.2020.148213\">https://doi.org/10.1016/j.bbabio.2020.148213</a>.","apa":"Adjobo-Hermans, M. J. W., De Haas, R., Willems, P. H. G. M., Wojtala, A., Van Emst-De Vries, S. E., Wagenaars, J. A., … Koopman, W. J. H. (2020). NDUFS4 deletion triggers loss of NDUFA12 in Ndufs4−/− mice and Leigh syndrome patients: A stabilizing role for NDUFAF2. <i>Biochimica et Biophysica Acta - Bioenergetics</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.bbabio.2020.148213\">https://doi.org/10.1016/j.bbabio.2020.148213</a>","mla":"Adjobo-Hermans, Merel J. W., et al. “NDUFS4 Deletion Triggers Loss of NDUFA12 in Ndufs4−/− Mice and Leigh Syndrome Patients: A Stabilizing Role for NDUFAF2.” <i>Biochimica et Biophysica Acta - Bioenergetics</i>, vol. 1861, no. 8, 148213, Elsevier, 2020, doi:<a href=\"https://doi.org/10.1016/j.bbabio.2020.148213\">10.1016/j.bbabio.2020.148213</a>.","ama":"Adjobo-Hermans MJW, De Haas R, Willems PHGM, et al. NDUFS4 deletion triggers loss of NDUFA12 in Ndufs4−/− mice and Leigh syndrome patients: A stabilizing role for NDUFAF2. <i>Biochimica et Biophysica Acta - Bioenergetics</i>. 2020;1861(8). doi:<a href=\"https://doi.org/10.1016/j.bbabio.2020.148213\">10.1016/j.bbabio.2020.148213</a>","ieee":"M. J. W. Adjobo-Hermans <i>et al.</i>, “NDUFS4 deletion triggers loss of NDUFA12 in Ndufs4−/− mice and Leigh syndrome patients: A stabilizing role for NDUFAF2,” <i>Biochimica et Biophysica Acta - Bioenergetics</i>, vol. 1861, no. 8. Elsevier, 2020.","short":"M.J.W. Adjobo-Hermans, R. De Haas, P.H.G.M. Willems, A. Wojtala, S.E. Van Emst-De Vries, J.A. Wagenaars, M. Van Den Brand, R.J. Rodenburg, J.A.M. Smeitink, L.G. Nijtmans, L.A. Sazanov, M.R. Wieckowski, W.J.H. Koopman, Biochimica et Biophysica Acta - Bioenergetics 1861 (2020)."},"language":[{"iso":"eng"}],"oa":1,"article_type":"original","date_created":"2020-05-03T22:00:47Z","volume":1861,"oa_version":"Published Version","title":"NDUFS4 deletion triggers loss of NDUFA12 in Ndufs4−/− mice and Leigh syndrome patients: A stabilizing role for NDUFAF2","author":[{"first_name":"Merel J.W.","full_name":"Adjobo-Hermans, Merel J.W.","last_name":"Adjobo-Hermans"},{"full_name":"De Haas, Ria","last_name":"De Haas","first_name":"Ria"},{"first_name":"Peter H.G.M.","full_name":"Willems, Peter H.G.M.","last_name":"Willems"},{"first_name":"Aleksandra","full_name":"Wojtala, Aleksandra","last_name":"Wojtala"},{"full_name":"Van Emst-De Vries, Sjenet E.","last_name":"Van Emst-De Vries","first_name":"Sjenet E."},{"full_name":"Wagenaars, Jori A.","last_name":"Wagenaars","first_name":"Jori A."},{"first_name":"Mariel","full_name":"Van Den Brand, Mariel","last_name":"Van Den Brand"},{"first_name":"Richard J.","last_name":"Rodenburg","full_name":"Rodenburg, Richard J."},{"first_name":"Jan A.M.","last_name":"Smeitink","full_name":"Smeitink, Jan A.M."},{"full_name":"Nijtmans, Leo G.","last_name":"Nijtmans","first_name":"Leo G."},{"first_name":"Leonid A","orcid":"0000-0002-0977-7989","last_name":"Sazanov","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A"},{"full_name":"Wieckowski, Mariusz R.","last_name":"Wieckowski","first_name":"Mariusz R."},{"full_name":"Koopman, Werner J.H.","last_name":"Koopman","first_name":"Werner J.H."}],"day":"01","scopus_import":"1","publication_status":"published","publication_identifier":{"eissn":["18792650"],"issn":["00052728"]},"file_date_updated":"2020-07-14T12:48:03Z","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"abstract":[{"lang":"eng","text":"Mutations in NDUFS4, which encodes an accessory subunit of mitochondrial oxidative phosphorylation (OXPHOS) complex I (CI), induce Leigh syndrome (LS). LS is a poorly understood pediatric disorder featuring brain-specific anomalies and early death. To study the LS pathomechanism, we here compared OXPHOS proteomes between various Ndufs4−/− mouse tissues. Ndufs4−/− animals displayed significantly lower CI subunit levels in brain/diaphragm relative to other tissues (liver/heart/kidney/skeletal muscle), whereas other OXPHOS subunit levels were not reduced. Absence of NDUFS4 induced near complete absence of the NDUFA12 accessory subunit, a 50% reduction in other CI subunit levels, and an increase in specific CI assembly factors. Among the latter, NDUFAF2 was most highly increased. Regarding NDUFS4, NDUFA12 and NDUFAF2, identical results were obtained in Ndufs4−/− mouse embryonic fibroblasts (MEFs) and NDUFS4-mutated LS patient cells. Ndufs4−/− MEFs contained active CI in situ but blue-native-PAGE highlighted that NDUFAF2 attached to an inactive CI subcomplex (CI-830) and inactive assemblies of higher MW. In NDUFA12-mutated LS patient cells, NDUFA12 absence did not reduce NDUFS4 levels but triggered NDUFAF2 association to active CI. BN-PAGE revealed no such association in LS patient fibroblasts with mutations in other CI subunit-encoding genes where NDUFAF2 was attached to CI-830 (NDUFS1, NDUFV1 mutation) or not detected (NDUFS7 mutation). Supported by enzymological and CI in silico structural analysis, we conclude that absence of NDUFS4 induces near complete absence of NDUFA12 but not vice versa, and that NDUFAF2 stabilizes active CI in Ndufs4−/− mice and LS patient cells, perhaps in concert with mitochondrial inner membrane lipids."}],"intvolume":"      1861","has_accepted_license":"1"},{"publisher":"American Chemical Society","article_processing_charge":"No","doi":"10.1021/jacs.9b13450","type":"journal_article","_id":"8040","date_updated":"2023-08-22T07:49:38Z","page":"9220-9230","quality_controlled":"1","related_material":{"record":[{"status":"public","relation":"research_data","id":"9326"},{"status":"public","relation":"research_data","id":"9713"},{"status":"public","relation":"research_data","id":"9878"}]},"external_id":{"isi":["000537415600020"],"pmid":["32347721"]},"year":"2020","isi":1,"status":"public","publication":"Journal of the American Chemical Society","date_published":"2020-05-20T00:00:00Z","pmid":1,"oa_version":"None","title":"Charge transfer and chemo-mechanical coupling in respiratory complex I","day":"20","scopus_import":"1","author":[{"first_name":"Chitrak","last_name":"Gupta","full_name":"Gupta, Chitrak"},{"first_name":"Umesh","last_name":"Khaniya","full_name":"Khaniya, Umesh"},{"full_name":"Chan, Chun Kit","last_name":"Chan","first_name":"Chun Kit"},{"full_name":"Dehez, Francois","last_name":"Dehez","first_name":"Francois"},{"first_name":"Mrinal","last_name":"Shekhar","full_name":"Shekhar, Mrinal"},{"full_name":"Gunner, M. R.","last_name":"Gunner","first_name":"M. R."},{"full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989"},{"full_name":"Chipot, Christophe","last_name":"Chipot","first_name":"Christophe"},{"last_name":"Singharoy","full_name":"Singharoy, Abhishek","first_name":"Abhishek"}],"date_created":"2020-06-29T07:59:35Z","article_type":"original","volume":142,"abstract":[{"text":"The mitochondrial respiratory chain, formed by five protein complexes, utilizes energy from catabolic processes to synthesize ATP. Complex I, the first and the largest protein complex of the chain, harvests electrons from NADH to reduce quinone, while pumping protons across the mitochondrial membrane. Detailed knowledge of the working principle of such coupled charge-transfer processes remains, however, fragmentary due to bottlenecks in understanding redox-driven conformational transitions and their interplay with the hydrated proton pathways. Complex I from Thermus thermophilus encases 16 subunits with nine iron–sulfur clusters, reduced by electrons from NADH. Here, employing the latest crystal structure of T. thermophilus complex I, we have used microsecond-scale molecular dynamics simulations to study the chemo-mechanical coupling between redox changes of the iron–sulfur clusters and conformational transitions across complex I. First, we identify the redox switches within complex I, which allosterically couple the dynamics of the quinone binding pocket to the site of NADH reduction. Second, our free-energy calculations reveal that the affinity of the quinone, specifically menaquinone, for the binding-site is higher than that of its reduced, menaquinol form—a design essential for menaquinol release. Remarkably, the barriers to diffusive menaquinone dynamics are lesser than that of the more ubiquitous ubiquinone, and the naphthoquinone headgroup of the former furnishes stronger binding interactions with the pocket, favoring menaquinone for charge transport in T. thermophilus. Our computations are consistent with experimentally validated mutations and hierarchize the key residues into three functional classes, identifying new mutation targets. Third, long-range hydrogen-bond networks connecting the quinone-binding site to the transmembrane subunits are found to be responsible for proton pumping. Put together, the simulations reveal the molecular design principles linking redox reactions to quinone turnover to proton translocation in complex I.","lang":"eng"}],"intvolume":"       142","publication_status":"published","publication_identifier":{"issn":["00027863"],"eissn":["15205126"]},"month":"05","department":[{"_id":"LeSa"}],"language":[{"iso":"eng"}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"ama":"Gupta C, Khaniya U, Chan CK, et al. Charge transfer and chemo-mechanical coupling in respiratory complex I. <i>Journal of the American Chemical Society</i>. 2020;142(20):9220-9230. doi:<a href=\"https://doi.org/10.1021/jacs.9b13450\">10.1021/jacs.9b13450</a>","short":"C. Gupta, U. Khaniya, C.K. Chan, F. Dehez, M. Shekhar, M.R. Gunner, L.A. Sazanov, C. Chipot, A. Singharoy, Journal of the American Chemical Society 142 (2020) 9220–9230.","ieee":"C. Gupta <i>et al.</i>, “Charge transfer and chemo-mechanical coupling in respiratory complex I,” <i>Journal of the American Chemical Society</i>, vol. 142, no. 20. American Chemical Society, pp. 9220–9230, 2020.","chicago":"Gupta, Chitrak, Umesh Khaniya, Chun Kit Chan, Francois Dehez, Mrinal Shekhar, M. R. Gunner, Leonid A Sazanov, Christophe Chipot, and Abhishek Singharoy. “Charge Transfer and Chemo-Mechanical Coupling in Respiratory Complex I.” <i>Journal of the American Chemical Society</i>. American Chemical Society, 2020. <a href=\"https://doi.org/10.1021/jacs.9b13450\">https://doi.org/10.1021/jacs.9b13450</a>.","ista":"Gupta C, Khaniya U, Chan CK, Dehez F, Shekhar M, Gunner MR, Sazanov LA, Chipot C, Singharoy A. 2020. Charge transfer and chemo-mechanical coupling in respiratory complex I. Journal of the American Chemical Society. 142(20), 9220–9230.","mla":"Gupta, Chitrak, et al. “Charge Transfer and Chemo-Mechanical Coupling in Respiratory Complex I.” <i>Journal of the American Chemical Society</i>, vol. 142, no. 20, American Chemical Society, 2020, pp. 9220–30, doi:<a href=\"https://doi.org/10.1021/jacs.9b13450\">10.1021/jacs.9b13450</a>.","apa":"Gupta, C., Khaniya, U., Chan, C. K., Dehez, F., Shekhar, M., Gunner, M. R., … Singharoy, A. (2020). Charge transfer and chemo-mechanical coupling in respiratory complex I. <i>Journal of the American Chemical Society</i>. American Chemical Society. <a href=\"https://doi.org/10.1021/jacs.9b13450\">https://doi.org/10.1021/jacs.9b13450</a>"},"issue":"20"},{"month":"07","department":[{"_id":"LeSa"}],"file":[{"file_size":7320493,"date_created":"2020-08-24T13:31:53Z","date_updated":"2020-08-24T13:31:53Z","creator":"cziletti","file_id":"8289","success":1,"file_name":"2020_eLife_Steiner.pdf","access_level":"open_access","content_type":"application/pdf","relation":"main_file","checksum":"b3656d14d5ddbb9d26e3074eea2d0c15"}],"article_number":"e59407","oa":1,"language":[{"iso":"eng"}],"citation":{"short":"J. Steiner, L.A. Sazanov, ELife 9 (2020).","ieee":"J. Steiner and L. A. Sazanov, “Structure and mechanism of the Mrp complex, an ancient cation/proton antiporter,” <i>eLife</i>, vol. 9. eLife Sciences Publications, 2020.","ama":"Steiner J, Sazanov LA. Structure and mechanism of the Mrp complex, an ancient cation/proton antiporter. <i>eLife</i>. 2020;9. doi:<a href=\"https://doi.org/10.7554/eLife.59407\">10.7554/eLife.59407</a>","mla":"Steiner, Julia, and Leonid A. Sazanov. “Structure and Mechanism of the Mrp Complex, an Ancient Cation/Proton Antiporter.” <i>ELife</i>, vol. 9, e59407, eLife Sciences Publications, 2020, doi:<a href=\"https://doi.org/10.7554/eLife.59407\">10.7554/eLife.59407</a>.","apa":"Steiner, J., &#38; Sazanov, L. A. (2020). Structure and mechanism of the Mrp complex, an ancient cation/proton antiporter. <i>ELife</i>. eLife Sciences Publications. <a href=\"https://doi.org/10.7554/eLife.59407\">https://doi.org/10.7554/eLife.59407</a>","chicago":"Steiner, Julia, and Leonid A Sazanov. “Structure and Mechanism of the Mrp Complex, an Ancient Cation/Proton Antiporter.” <i>ELife</i>. eLife Sciences Publications, 2020. <a href=\"https://doi.org/10.7554/eLife.59407\">https://doi.org/10.7554/eLife.59407</a>.","ista":"Steiner J, Sazanov LA. 2020. Structure and mechanism of the Mrp complex, an ancient cation/proton antiporter. eLife. 9, e59407."},"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","author":[{"orcid":"0000-0003-0493-3775","first_name":"Julia","id":"3BB67EB0-F248-11E8-B48F-1D18A9856A87","full_name":"Steiner, Julia","last_name":"Steiner"},{"orcid":"0000-0002-0977-7989","first_name":"Leonid A","last_name":"Sazanov","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A"}],"scopus_import":"1","day":"31","title":"Structure and mechanism of the Mrp complex, an ancient cation/proton antiporter","oa_version":"Published Version","volume":9,"article_type":"original","date_created":"2020-08-24T06:24:04Z","has_accepted_license":"1","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"intvolume":"         9","abstract":[{"lang":"eng","text":"Multiple resistance and pH adaptation (Mrp) antiporters are multi-subunit Na+ (or K+)/H+ exchangers representing an ancestor of many essential redox-driven proton pumps, such as respiratory complex I. The mechanism of coupling between ion or electron transfer and proton translocation in this large protein family is unknown. Here, we present the structure of the Mrp complex from Anoxybacillus flavithermus solved by cryo-EM at 3.0 Å resolution. It is a dimer of seven-subunit protomers with 50 trans-membrane helices each. Surface charge distribution within each monomer is remarkably asymmetric, revealing probable proton and sodium translocation pathways. On the basis of the structure we propose a mechanism where the coupling between sodium and proton translocation is facilitated by a series of electrostatic interactions between a cation and key charged residues. This mechanism is likely to be applicable to the entire family of redox proton pumps, where electron transfer to substrates replaces cation movements."}],"acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"LifeSc"}],"publication_status":"published","publication_identifier":{"eissn":["2050084X"]},"file_date_updated":"2020-08-24T13:31:53Z","isi":1,"year":"2020","external_id":{"pmid":["32735215"],"isi":["000562123600001"]},"related_material":{"record":[{"status":"public","relation":"dissertation_contains","id":"8353"}],"link":[{"description":"News on IST Homepage","url":"https://ist.ac.at/en/news/mystery-of-giant-proton-pump-solved/","relation":"press_release"}]},"project":[{"_id":"26169496-B435-11E9-9278-68D0E5697425","grant_number":"24741","name":"Revealing the functional mechanism of Mrp antiporter, an ancestor of complex I"}],"publication":"eLife","status":"public","pmid":1,"date_published":"2020-07-31T00:00:00Z","acknowledgement":"This research was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Electron Microscopy Facility (EMF), the Life Science Facility (LSF) and the IST high-performance computing cluster. We thank Dr Victor-Valentin Hodirnau and Daniel Johann Gütl from IST Austria for assistance with collecting cryo-EM data. We thank Prof. Masahiro Ito (Graduate School of Life Sciences, Toyo University, Japan) for a kind provision of plasmid DNA encoding Mrp from A. flavithermus WK1. JS is a recipient of a DOC Fellowship of the Austrian Academy of Sciences at the Institute of Science and Technology, Austria.","doi":"10.7554/eLife.59407","article_processing_charge":"No","publisher":"eLife Sciences Publications","date_updated":"2023-09-07T13:14:08Z","_id":"8284","type":"journal_article","ddc":["570"],"quality_controlled":"1"},{"oa_version":"Published Version","title":"Key role of quinone in the mechanism of respiratory complex I","author":[{"first_name":"Javier","full_name":"Gutierrez-Fernandez, Javier","id":"3D9511BA-F248-11E8-B48F-1D18A9856A87","last_name":"Gutierrez-Fernandez"},{"first_name":"Karol","last_name":"Kaszuba","full_name":"Kaszuba, Karol","id":"3FDF9472-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Minhas","full_name":"Minhas, Gurdeep S.","first_name":"Gurdeep S."},{"last_name":"Baradaran","full_name":"Baradaran, Rozbeh","first_name":"Rozbeh"},{"full_name":"Tambalo, Margherita","id":"4187dfe4-ec23-11ea-ae46-f08ab378313a","last_name":"Tambalo","first_name":"Margherita"},{"first_name":"David T.","full_name":"Gallagher, David T.","last_name":"Gallagher"},{"first_name":"Leonid A","orcid":"0000-0002-0977-7989","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","last_name":"Sazanov"}],"scopus_import":"1","day":"18","article_type":"original","date_created":"2020-08-30T22:01:10Z","volume":11,"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","short":"CC BY (4.0)"},"intvolume":"        11","abstract":[{"lang":"eng","text":"Complex I is the first and the largest enzyme of respiratory chains in bacteria and mitochondria. The mechanism which couples spatially separated transfer of electrons to proton translocation in complex I is not known. Here we report five crystal structures of T. thermophilus enzyme in complex with NADH or quinone-like compounds. We also determined cryo-EM structures of major and minor native states of the complex, differing in the position of the peripheral arm. Crystal structures show that binding of quinone-like compounds (but not of NADH) leads to a related global conformational change, accompanied by local re-arrangements propagating from the quinone site to the nearest proton channel. Normal mode and molecular dynamics analyses indicate that these are likely to represent the first steps in the proton translocation mechanism. Our results suggest that quinone binding and chemistry play a key role in the coupling mechanism of complex I."}],"has_accepted_license":"1","publication_status":"published","publication_identifier":{"eissn":["20411723"]},"file_date_updated":"2020-08-31T13:40:00Z","month":"08","file":[{"file_id":"8326","creator":"cziletti","date_updated":"2020-08-31T13:40:00Z","date_created":"2020-08-31T13:40:00Z","file_size":7527373,"checksum":"52b96f41d7d0db9728064c08da00d030","relation":"main_file","content_type":"application/pdf","access_level":"open_access","file_name":"2020_NatComm_Gutierrez-Fernandez.pdf","success":1}],"article_number":"4135","department":[{"_id":"LeSa"}],"language":[{"iso":"eng"}],"oa":1,"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","issue":"1","citation":{"short":"J. Gutierrez-Fernandez, K. Kaszuba, G.S. Minhas, R. Baradaran, M. Tambalo, D.T. Gallagher, L.A. Sazanov, Nature Communications 11 (2020).","ieee":"J. Gutierrez-Fernandez <i>et al.</i>, “Key role of quinone in the mechanism of respiratory complex I,” <i>Nature Communications</i>, vol. 11, no. 1. Springer Nature, 2020.","ama":"Gutierrez-Fernandez J, Kaszuba K, Minhas GS, et al. Key role of quinone in the mechanism of respiratory complex I. <i>Nature Communications</i>. 2020;11(1). doi:<a href=\"https://doi.org/10.1038/s41467-020-17957-0\">10.1038/s41467-020-17957-0</a>","mla":"Gutierrez-Fernandez, Javier, et al. “Key Role of Quinone in the Mechanism of Respiratory Complex I.” <i>Nature Communications</i>, vol. 11, no. 1, 4135, Springer Nature, 2020, doi:<a href=\"https://doi.org/10.1038/s41467-020-17957-0\">10.1038/s41467-020-17957-0</a>.","apa":"Gutierrez-Fernandez, J., Kaszuba, K., Minhas, G. S., Baradaran, R., Tambalo, M., Gallagher, D. T., &#38; Sazanov, L. A. (2020). Key role of quinone in the mechanism of respiratory complex I. <i>Nature Communications</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41467-020-17957-0\">https://doi.org/10.1038/s41467-020-17957-0</a>","ista":"Gutierrez-Fernandez J, Kaszuba K, Minhas GS, Baradaran R, Tambalo M, Gallagher DT, Sazanov LA. 2020. Key role of quinone in the mechanism of respiratory complex I. Nature Communications. 11(1), 4135.","chicago":"Gutierrez-Fernandez, Javier, Karol Kaszuba, Gurdeep S. Minhas, Rozbeh Baradaran, Margherita Tambalo, David T. Gallagher, and Leonid A Sazanov. “Key Role of Quinone in the Mechanism of Respiratory Complex I.” <i>Nature Communications</i>. Springer Nature, 2020. <a href=\"https://doi.org/10.1038/s41467-020-17957-0\">https://doi.org/10.1038/s41467-020-17957-0</a>."},"publisher":"Springer Nature","doi":"10.1038/s41467-020-17957-0","article_processing_charge":"No","type":"journal_article","date_updated":"2023-08-22T09:03:00Z","_id":"8318","ddc":["570"],"quality_controlled":"1","external_id":{"isi":["000607072900001"],"pmid":["32811817"]},"related_material":{"link":[{"description":"News on IST Homepage","url":"https://ist.ac.at/en/news/mystery-of-giant-proton-pump-solved/","relation":"press_release"}]},"isi":1,"year":"2020","publication":"Nature Communications","status":"public","acknowledgement":"This work was funded by the Medical Research Council, UK and IST Austria. We thank the European Synchrotron Radiation Facility and the Diamond Light Source for provision of synchrotron radiation facilities. We are grateful to the staff of beamlines ID29, ID23-2 (ESRF, Grenoble, France) and I03 (Diamond Light Source, Didcot, UK) for assistance. Data processing was performed at the IST high-performance computing cluster.","date_published":"2020-08-18T00:00:00Z","pmid":1},{"year":"2020","isi":1,"related_material":{"link":[{"description":"News on IST Homepage","url":"https://ist.ac.at/en/news/structure-of-atpase-solved/","relation":"press_release"}]},"external_id":{"isi":["000569299400004"],"pmid":["32929284"]},"publication":"Nature Structural and Molecular Biology","status":"public","pmid":1,"acknowledgement":"We thank J. Novacek from CEITEC (Brno, Czech Republic) for assistance with collecting the FEI Krios dataset and iNEXT for providing access to CEITEC. We thank the IST Austria EM facility for access and assistance with collecting the FEI Glacios dataset. Data processing was performed at the IST high-performance computing cluster. This work has been supported by iNEXT EM HEDC (proposal 4506), funded by the Horizon 2020 Programme of the European Commission.","date_published":"2020-11-01T00:00:00Z","article_processing_charge":"No","doi":"10.1038/s41594-020-0503-8","publisher":"Springer Nature","_id":"8581","date_updated":"2023-08-22T09:33:09Z","type":"journal_article","page":"1077-1085","quality_controlled":"1","month":"11","department":[{"_id":"LeSa"}],"language":[{"iso":"eng"}],"citation":{"ama":"Pinke G, Zhou L, Sazanov LA. Cryo-EM structure of the entire mammalian F-type ATP synthase. <i>Nature Structural and Molecular Biology</i>. 2020;27(11):1077-1085. doi:<a href=\"https://doi.org/10.1038/s41594-020-0503-8\">10.1038/s41594-020-0503-8</a>","ieee":"G. Pinke, L. Zhou, and L. A. Sazanov, “Cryo-EM structure of the entire mammalian F-type ATP synthase,” <i>Nature Structural and Molecular Biology</i>, vol. 27, no. 11. Springer Nature, pp. 1077–1085, 2020.","short":"G. Pinke, L. Zhou, L.A. Sazanov, Nature Structural and Molecular Biology 27 (2020) 1077–1085.","ista":"Pinke G, Zhou L, Sazanov LA. 2020. Cryo-EM structure of the entire mammalian F-type ATP synthase. Nature Structural and Molecular Biology. 27(11), 1077–1085.","chicago":"Pinke, Gergely, Long Zhou, and Leonid A Sazanov. “Cryo-EM Structure of the Entire Mammalian F-Type ATP Synthase.” <i>Nature Structural and Molecular Biology</i>. Springer Nature, 2020. <a href=\"https://doi.org/10.1038/s41594-020-0503-8\">https://doi.org/10.1038/s41594-020-0503-8</a>.","apa":"Pinke, G., Zhou, L., &#38; Sazanov, L. A. (2020). Cryo-EM structure of the entire mammalian F-type ATP synthase. <i>Nature Structural and Molecular Biology</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41594-020-0503-8\">https://doi.org/10.1038/s41594-020-0503-8</a>","mla":"Pinke, Gergely, et al. “Cryo-EM Structure of the Entire Mammalian F-Type ATP Synthase.” <i>Nature Structural and Molecular Biology</i>, vol. 27, no. 11, Springer Nature, 2020, pp. 1077–85, doi:<a href=\"https://doi.org/10.1038/s41594-020-0503-8\">10.1038/s41594-020-0503-8</a>."},"issue":"11","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","day":"01","scopus_import":"1","author":[{"last_name":"Pinke","id":"4D5303E6-F248-11E8-B48F-1D18A9856A87","full_name":"Pinke, Gergely","first_name":"Gergely"},{"full_name":"Zhou, Long","id":"3E751364-F248-11E8-B48F-1D18A9856A87","last_name":"Zhou","orcid":"0000-0002-1864-8951","first_name":"Long"},{"last_name":"Sazanov","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","first_name":"Leonid A","orcid":"0000-0002-0977-7989"}],"oa_version":"None","title":"Cryo-EM structure of the entire mammalian F-type ATP synthase","volume":27,"date_created":"2020-09-28T08:59:27Z","article_type":"original","acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"ScienComp"}],"abstract":[{"text":"The majority of adenosine triphosphate (ATP) powering cellular processes in eukaryotes is produced by the mitochondrial F1Fo ATP synthase. Here, we present the atomic models of the membrane Fo domain and the entire mammalian (ovine) F1Fo, determined by cryo-electron microscopy. Subunits in the membrane domain are arranged in the ‘proton translocation cluster’ attached to the c-ring and a more distant ‘hook apparatus’ holding subunit e. Unexpectedly, this subunit is anchored to a lipid ‘plug’ capping the c-ring. We present a detailed proton translocation pathway in mammalian Fo and key inter-monomer contacts in F1Fo multimers. Cryo-EM maps of F1Fo exposed to calcium reveal a retracted subunit e and a disassembled c-ring, suggesting permeability transition pore opening. We propose a model for the permeability transition pore opening, whereby subunit e pulls the lipid plug out of the c-ring. Our structure will allow the design of drugs for many emerging applications in medicine.","lang":"eng"}],"intvolume":"        27","publication_identifier":{"issn":["15459993"],"eissn":["15459985"]},"publication_status":"published"},{"ddc":["572"],"quality_controlled":"1","article_processing_charge":"No","doi":"10.1126/science.abc4209","publisher":"American Association for the Advancement of Science","_id":"8737","date_updated":"2023-08-22T12:35:38Z","type":"journal_article","status":"public","publication":"Science","project":[{"call_identifier":"H2020","grant_number":"665385","name":"International IST Doctoral Program","_id":"2564DBCA-B435-11E9-9278-68D0E5697425"}],"ec_funded":1,"pmid":1,"acknowledgement":"We thank J. Novacek (CEITEC Brno) and V.-V. Hodirnau (IST Austria) for their help with collecting cryo-EM datasets. We thank the IST Life Science and Electron Microscopy Facilities for providing equipment. This work has been supported by iNEXT,project number 653706, funded by the Horizon 2020 program of the European Union. This article reflects only the authors’view,and the European Commission is not responsible for any use that may be made of the information it contains. CIISB research infrastructure project LM2015043 funded by MEYS CR is gratefully acknowledged for the financial support of the measurements at the CF Cryo-electron Microscopy and Tomography CEITEC MU.This project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement no. 665385","date_published":"2020-10-30T00:00:00Z","year":"2020","isi":1,"external_id":{"isi":["000583031800004"],"pmid":["32972993"]},"has_accepted_license":"1","acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"EM-Fac"}],"abstract":[{"text":"Mitochondrial complex I couples NADH:ubiquinone oxidoreduction to proton pumping by an unknown mechanism. Here, we present cryo-electron microscopy structures of ovine complex I in five different conditions, including turnover, at resolutions up to 2.3 to 2.5 angstroms. Resolved water molecules allowed us to experimentally define the proton translocation pathways. Quinone binds at three positions along the quinone cavity, as does the inhibitor rotenone that also binds within subunit ND4. Dramatic conformational changes around the quinone cavity couple the redox reaction to proton translocation during open-to-closed state transitions of the enzyme. In the induced deactive state, the open conformation is arrested by the ND6 subunit. We propose a detailed molecular coupling mechanism of complex I, which is an unexpected combination of conformational changes and electrostatic interactions.","lang":"eng"}],"intvolume":"       370","file_date_updated":"2020-11-26T18:47:58Z","publication_identifier":{"eissn":["10959203"]},"publication_status":"published","scopus_import":"1","day":"30","author":[{"first_name":"Domen","last_name":"Kampjut","full_name":"Kampjut, Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989"}],"title":"The coupling mechanism of mammalian respiratory complex I","oa_version":"Submitted Version","volume":370,"date_created":"2020-11-08T23:01:23Z","article_type":"original","oa":1,"language":[{"iso":"eng"}],"citation":{"short":"D. Kampjut, L.A. Sazanov, Science 370 (2020).","ieee":"D. Kampjut and L. A. Sazanov, “The coupling mechanism of mammalian respiratory complex I,” <i>Science</i>, vol. 370, no. 6516. American Association for the Advancement of Science, 2020.","ama":"Kampjut D, Sazanov LA. The coupling mechanism of mammalian respiratory complex I. <i>Science</i>. 2020;370(6516). doi:<a href=\"https://doi.org/10.1126/science.abc4209\">10.1126/science.abc4209</a>","mla":"Kampjut, Domen, and Leonid A. Sazanov. “The Coupling Mechanism of Mammalian Respiratory Complex I.” <i>Science</i>, vol. 370, no. 6516, eabc4209, American Association for the Advancement of Science, 2020, doi:<a href=\"https://doi.org/10.1126/science.abc4209\">10.1126/science.abc4209</a>.","apa":"Kampjut, D., &#38; Sazanov, L. A. (2020). The coupling mechanism of mammalian respiratory complex I. <i>Science</i>. American Association for the Advancement of Science. <a href=\"https://doi.org/10.1126/science.abc4209\">https://doi.org/10.1126/science.abc4209</a>","chicago":"Kampjut, Domen, and Leonid A Sazanov. “The Coupling Mechanism of Mammalian Respiratory Complex I.” <i>Science</i>. American Association for the Advancement of Science, 2020. <a href=\"https://doi.org/10.1126/science.abc4209\">https://doi.org/10.1126/science.abc4209</a>.","ista":"Kampjut D, Sazanov LA. 2020. The coupling mechanism of mammalian respiratory complex I. Science. 370(6516), eabc4209."},"issue":"6516","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","month":"10","department":[{"_id":"LeSa"}],"article_number":"eabc4209","file":[{"file_id":"8820","date_updated":"2020-11-26T18:47:58Z","creator":"lsazanov","file_size":7618987,"date_created":"2020-11-26T18:47:58Z","checksum":"658ba90979ca9528a2efdfac8547047a","relation":"main_file","access_level":"open_access","content_type":"application/pdf","success":1,"file_name":"Full_manuscript_with_SI_opt_red.pdf"}]},{"citation":{"ieee":"C. Gupta <i>et al.</i>, “Charge transfer and chemo-mechanical coupling in respiratory complex I.” American Chemical Society, 2020.","short":"C. Gupta, U. Khaniya, C. Chan, F. Dehez, M. Shekhar, M.R. Gunner, L.A. Sazanov, C. Chipot, A. Singharoy, (2020).","ama":"Gupta C, Khaniya U, Chan C, et al. Charge transfer and chemo-mechanical coupling in respiratory complex I. 2020. doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>","apa":"Gupta, C., Khaniya, U., Chan, C., Dehez, F., Shekhar, M., Gunner, M. R., … Singharoy, A. (2020). Charge transfer and chemo-mechanical coupling in respiratory complex I. American Chemical Society. <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">https://doi.org/10.1021/jacs.9b13450.s002</a>","mla":"Gupta, Chitrak, et al. <i>Charge Transfer and Chemo-Mechanical Coupling in Respiratory Complex I</i>. American Chemical Society, 2020, doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>.","ista":"Gupta C, Khaniya U, Chan C, Dehez F, Shekhar M, Gunner MR, Sazanov LA, Chipot C, Singharoy A. 2020. Charge transfer and chemo-mechanical coupling in respiratory complex I, American Chemical Society, <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>.","chicago":"Gupta, Chitrak, Umesh Khaniya, Chun Chan, Francois Dehez, Mrinal Shekhar, M. R. Gunner, Leonid A Sazanov, Christophe Chipot, and Abhishek Singharoy. “Charge Transfer and Chemo-Mechanical Coupling in Respiratory Complex I.” American Chemical Society, 2020. <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">https://doi.org/10.1021/jacs.9b13450.s002</a>."},"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","date_published":"2020-05-20T00:00:00Z","oa":1,"status":"public","department":[{"_id":"LeSa"}],"year":"2020","related_material":{"record":[{"status":"public","relation":"used_in_publication","id":"8040"}]},"month":"05","main_file_link":[{"open_access":"1"}],"license":"https://creativecommons.org/licenses/by-nc/4.0/","tmp":{"name":"Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc/4.0/legalcode","image":"/images/cc_by_nc.png","short":"CC BY-NC (4.0)"},"abstract":[{"text":"The mitochondrial respiratory chain, formed by five protein complexes, utilizes energy from catabolic processes to synthesize ATP. Complex I, the first and the largest protein complex of the chain, harvests electrons from NADH to reduce quinone, while pumping protons across the mitochondrial membrane. Detailed knowledge of the working principle of such coupled charge-transfer processes remains, however, fragmentary due to bottlenecks in understanding redox-driven conformational transitions and their interplay with the hydrated proton pathways. Complex I from Thermus thermophilus encases 16 subunits with nine iron–sulfur clusters, reduced by electrons from NADH. Here, employing the latest crystal structure of T. thermophilus complex I, we have used microsecond-scale molecular dynamics simulations to study the chemo-mechanical coupling between redox changes of the iron–sulfur clusters and conformational transitions across complex I. First, we identify the redox switches within complex I, which allosterically couple the dynamics of the quinone binding pocket to the site of NADH reduction. Second, our free-energy calculations reveal that the affinity of the quinone, specifically menaquinone, for the binding-site is higher than that of its reduced, menaquinol forma design essential for menaquinol release. Remarkably, the barriers to diffusive menaquinone dynamics are lesser than that of the more ubiquitous ubiquinone, and the naphthoquinone headgroup of the former furnishes stronger binding interactions with the pocket, favoring menaquinone for charge transport in T. thermophilus. Our computations are consistent with experimentally validated mutations and hierarchize the key residues into three functional classes, identifying new mutation targets. Third, long-range hydrogen-bond networks connecting the quinone-binding site to the transmembrane subunits are found to be responsible for proton pumping. Put together, the simulations reveal the molecular design principles linking redox reactions to quinone turnover to proton translocation in complex I.","lang":"eng"}],"_id":"9326","date_updated":"2023-08-22T07:49:37Z","date_created":"2021-04-14T12:05:20Z","type":"research_data_reference","day":"20","article_processing_charge":"No","author":[{"full_name":"Gupta, Chitrak","last_name":"Gupta","first_name":"Chitrak"},{"last_name":"Khaniya","full_name":"Khaniya, Umesh","first_name":"Umesh"},{"first_name":"Chun","last_name":"Chan","full_name":"Chan, Chun"},{"first_name":"Francois","full_name":"Dehez, Francois","last_name":"Dehez"},{"first_name":"Mrinal","last_name":"Shekhar","full_name":"Shekhar, Mrinal"},{"first_name":"M. R.","last_name":"Gunner","full_name":"Gunner, M. R."},{"full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989"},{"first_name":"Christophe","last_name":"Chipot","full_name":"Chipot, Christophe"},{"first_name":"Abhishek","last_name":"Singharoy","full_name":"Singharoy, Abhishek"}],"doi":"10.1021/jacs.9b13450.s002","publisher":"American Chemical Society","oa_version":"Published Version","title":"Charge transfer and chemo-mechanical coupling in respiratory complex I"},{"abstract":[{"text":"Additional analyses of the trajectories","lang":"eng"}],"article_processing_charge":"No","day":"20","author":[{"full_name":"Gupta, Chitrak","last_name":"Gupta","first_name":"Chitrak"},{"first_name":"Umesh","last_name":"Khaniya","full_name":"Khaniya, Umesh"},{"first_name":"Chun Kit","last_name":"Chan","full_name":"Chan, Chun Kit"},{"last_name":"Dehez","full_name":"Dehez, Francois","first_name":"Francois"},{"first_name":"Mrinal","last_name":"Shekhar","full_name":"Shekhar, Mrinal"},{"last_name":"Gunner","full_name":"Gunner, M.R.","first_name":"M.R."},{"last_name":"Sazanov","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-0977-7989","first_name":"Leonid A"},{"last_name":"Chipot","full_name":"Chipot, Christophe","first_name":"Christophe"},{"full_name":"Singharoy, Abhishek","last_name":"Singharoy","first_name":"Abhishek"}],"doi":"10.1021/jacs.9b13450.s001","oa_version":"Published Version","title":"Supporting information","publisher":"American Chemical Society ","_id":"9713","date_updated":"2023-08-22T07:49:38Z","date_created":"2021-07-23T12:02:39Z","type":"research_data_reference","status":"public","citation":{"apa":"Gupta, C., Khaniya, U., Chan, C. K., Dehez, F., Shekhar, M., Gunner, M. R., … Singharoy, A. (2020). Supporting information. American Chemical Society . <a href=\"https://doi.org/10.1021/jacs.9b13450.s001\">https://doi.org/10.1021/jacs.9b13450.s001</a>","mla":"Gupta, Chitrak, et al. <i>Supporting Information</i>. American Chemical Society , 2020, doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s001\">10.1021/jacs.9b13450.s001</a>.","ista":"Gupta C, Khaniya U, Chan CK, Dehez F, Shekhar M, Gunner MR, Sazanov LA, Chipot C, Singharoy A. 2020. Supporting information, American Chemical Society , <a href=\"https://doi.org/10.1021/jacs.9b13450.s001\">10.1021/jacs.9b13450.s001</a>.","chicago":"Gupta, Chitrak, Umesh Khaniya, Chun Kit Chan, Francois Dehez, Mrinal Shekhar, M.R. Gunner, Leonid A Sazanov, Christophe Chipot, and Abhishek Singharoy. “Supporting Information.” American Chemical Society , 2020. <a href=\"https://doi.org/10.1021/jacs.9b13450.s001\">https://doi.org/10.1021/jacs.9b13450.s001</a>.","ieee":"C. Gupta <i>et al.</i>, “Supporting information.” American Chemical Society , 2020.","short":"C. Gupta, U. Khaniya, C.K. Chan, F. Dehez, M. Shekhar, M.R. Gunner, L.A. Sazanov, C. Chipot, A. Singharoy, (2020).","ama":"Gupta C, Khaniya U, Chan CK, et al. Supporting information. 2020. doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s001\">10.1021/jacs.9b13450.s001</a>"},"user_id":"6785fbc1-c503-11eb-8a32-93094b40e1cf","date_published":"2020-05-20T00:00:00Z","year":"2020","related_material":{"record":[{"id":"8040","relation":"used_in_publication","status":"public"}]},"month":"05","department":[{"_id":"LeSa"}]},{"author":[{"first_name":"Chitrak","last_name":"Gupta","full_name":"Gupta, Chitrak"},{"full_name":"Khaniya, Umesh","last_name":"Khaniya","first_name":"Umesh"},{"first_name":"Chun Kit","last_name":"Chan","full_name":"Chan, Chun Kit"},{"last_name":"Dehez","full_name":"Dehez, Francois","first_name":"Francois"},{"first_name":"Mrinal","last_name":"Shekhar","full_name":"Shekhar, Mrinal"},{"first_name":"M.R.","full_name":"Gunner, M.R.","last_name":"Gunner"},{"last_name":"Sazanov","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","orcid":"0000-0002-0977-7989","first_name":"Leonid A"},{"last_name":"Chipot","full_name":"Chipot, Christophe","first_name":"Christophe"},{"first_name":"Abhishek","full_name":"Singharoy, Abhishek","last_name":"Singharoy"}],"doi":"10.1021/jacs.9b13450.s002","article_processing_charge":"No","day":"20","title":"Movies","publisher":"American Chemical Society","oa_version":"Published Version","date_updated":"2023-08-22T07:49:38Z","_id":"9878","type":"research_data_reference","date_created":"2021-08-11T09:18:54Z","year":"2020","month":"05","related_material":{"record":[{"id":"8040","relation":"used_in_publication","status":"public"}]},"department":[{"_id":"LeSa"}],"status":"public","citation":{"chicago":"Gupta, Chitrak, Umesh Khaniya, Chun Kit Chan, Francois Dehez, Mrinal Shekhar, M.R. Gunner, Leonid A Sazanov, Christophe Chipot, and Abhishek Singharoy. “Movies.” American Chemical Society, 2020. <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">https://doi.org/10.1021/jacs.9b13450.s002</a>.","ista":"Gupta C, Khaniya U, Chan CK, Dehez F, Shekhar M, Gunner MR, Sazanov LA, Chipot C, Singharoy A. 2020. Movies, American Chemical Society, <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>.","mla":"Gupta, Chitrak, et al. <i>Movies</i>. American Chemical Society, 2020, doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>.","apa":"Gupta, C., Khaniya, U., Chan, C. K., Dehez, F., Shekhar, M., Gunner, M. R., … Singharoy, A. (2020). Movies. American Chemical Society. <a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">https://doi.org/10.1021/jacs.9b13450.s002</a>","ama":"Gupta C, Khaniya U, Chan CK, et al. Movies. 2020. doi:<a href=\"https://doi.org/10.1021/jacs.9b13450.s002\">10.1021/jacs.9b13450.s002</a>","short":"C. Gupta, U. Khaniya, C.K. Chan, F. Dehez, M. Shekhar, M.R. Gunner, L.A. Sazanov, C. Chipot, A. Singharoy, (2020).","ieee":"C. Gupta <i>et al.</i>, “Movies.” American Chemical Society, 2020."},"date_published":"2020-05-20T00:00:00Z","user_id":"6785fbc1-c503-11eb-8a32-93094b40e1cf"},{"related_material":{"link":[{"description":"News on IST Website","url":"https://ist.ac.at/en/news/high-end-microscopy-reveals-structure-and-function-of-crucial-metabolic-enzyme/","relation":"press_release"}],"record":[{"status":"public","relation":"dissertation_contains","id":"8340"}]},"external_id":{"pmid":["31462775"],"isi":["000485415400061"]},"year":"2019","isi":1,"status":"public","publication":"Nature","project":[{"_id":"2564DBCA-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","name":"International IST Doctoral Program","grant_number":"665385"}],"acknowledgement":" We thank R. Thompson, G. Effantin and V.-V. Hodirnau for their assistance with collecting NADP+, NADPH and apo datasets, respectively. Data processing was performed at the IST high-performance computing cluster.\r\nThis project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie Grant Agreement no. 665385.","date_published":"2019-09-12T00:00:00Z","ec_funded":1,"pmid":1,"publisher":"Springer Nature","article_processing_charge":"No","doi":"10.1038/s41586-019-1519-2","type":"journal_article","_id":"6848","date_updated":"2024-03-25T23:30:08Z","ddc":["572"],"page":"291–295","quality_controlled":"1","month":"09","file":[{"access_level":"open_access","content_type":"application/pdf","success":1,"file_name":"Manuscript_final_acc_withFigs_SI_opt_red.pdf","checksum":"52728cda5210a3e9b74cc204e8aed3d5","relation":"main_file","creator":"lsazanov","date_updated":"2020-11-26T16:33:44Z","date_created":"2020-11-26T16:33:44Z","file_size":3066206,"file_id":"8821"}],"department":[{"_id":"LeSa"}],"language":[{"iso":"eng"}],"oa":1,"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"apa":"Kampjut, D., &#38; Sazanov, L. A. (2019). Structure and mechanism of mitochondrial proton-translocating transhydrogenase. <i>Nature</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41586-019-1519-2\">https://doi.org/10.1038/s41586-019-1519-2</a>","mla":"Kampjut, Domen, and Leonid A. Sazanov. “Structure and Mechanism of Mitochondrial Proton-Translocating Transhydrogenase.” <i>Nature</i>, vol. 573, no. 7773, Springer Nature, 2019, pp. 291–295, doi:<a href=\"https://doi.org/10.1038/s41586-019-1519-2\">10.1038/s41586-019-1519-2</a>.","ista":"Kampjut D, Sazanov LA. 2019. Structure and mechanism of mitochondrial proton-translocating transhydrogenase. Nature. 573(7773), 291–295.","chicago":"Kampjut, Domen, and Leonid A Sazanov. “Structure and Mechanism of Mitochondrial Proton-Translocating Transhydrogenase.” <i>Nature</i>. Springer Nature, 2019. <a href=\"https://doi.org/10.1038/s41586-019-1519-2\">https://doi.org/10.1038/s41586-019-1519-2</a>.","ieee":"D. Kampjut and L. A. Sazanov, “Structure and mechanism of mitochondrial proton-translocating transhydrogenase,” <i>Nature</i>, vol. 573, no. 7773. Springer Nature, pp. 291–295, 2019.","short":"D. Kampjut, L.A. Sazanov, Nature 573 (2019) 291–295.","ama":"Kampjut D, Sazanov LA. Structure and mechanism of mitochondrial proton-translocating transhydrogenase. <i>Nature</i>. 2019;573(7773):291–295. doi:<a href=\"https://doi.org/10.1038/s41586-019-1519-2\">10.1038/s41586-019-1519-2</a>"},"issue":"7773","title":"Structure and mechanism of mitochondrial proton-translocating transhydrogenase","oa_version":"Submitted Version","scopus_import":"1","day":"12","author":[{"first_name":"Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87","full_name":"Kampjut, Domen","last_name":"Kampjut"},{"orcid":"0000-0002-0977-7989","first_name":"Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","last_name":"Sazanov"}],"date_created":"2019-09-04T06:21:41Z","article_type":"letter_note","volume":573,"acknowledged_ssus":[{"_id":"ScienComp"}],"abstract":[{"lang":"eng","text":"Proton-translocating transhydrogenase (also known as nicotinamide nucleotide transhydrogenase (NNT)) is found in the plasma membranes of bacteria and the inner mitochondrial membranes of eukaryotes. NNT catalyses the transfer of a hydride between NADH and NADP+, coupled to the translocation of one proton across the membrane. Its main physiological function is the generation of NADPH, which is a substrate in anabolic reactions and a regulator of oxidative status; however, NNT may also fine-tune the Krebs cycle1,2. NNT deficiency causes familial glucocorticoid deficiency in humans and metabolic abnormalities in mice, similar to those observed in type II diabetes3,4. The catalytic mechanism of NNT has been proposed to involve a rotation of around 180° of the entire NADP(H)-binding domain that alternately participates in hydride transfer and proton-channel gating. However, owing to the lack of high-resolution structures of intact NNT, the details of this process remain unclear5,6. Here we present the cryo-electron microscopy structure of intact mammalian NNT in different conformational states. We show how the NADP(H)-binding domain opens the proton channel to the opposite sides of the membrane, and we provide structures of these two states. We also describe the catalytically important interfaces and linkers between the membrane and the soluble domains and their roles in nucleotide exchange. These structures enable us to propose a revised mechanism for a coupling process in NNT that is consistent with a large body of previous biochemical work. Our results are relevant to the development of currently unavailable NNT inhibitors, which may have therapeutic potential in ischaemia reperfusion injury, metabolic syndrome and some cancers7,8,9."}],"intvolume":"       573","has_accepted_license":"1","file_date_updated":"2020-11-26T16:33:44Z","publication_status":"published","publication_identifier":{"issn":["0028-0836"],"eissn":["1476-4687"]}},{"language":[{"iso":"eng"}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"chicago":"Zhou, Long, and Leonid A Sazanov. “Structure and Conformational Plasticity of the Intact Thermus Thermophilus V/A-Type ATPase.” <i>Science</i>. AAAS, 2019. <a href=\"https://doi.org/10.1126/science.aaw9144\">https://doi.org/10.1126/science.aaw9144</a>.","ista":"Zhou L, Sazanov LA. 2019. Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase. Science. 365(6455), eaaw9144.","mla":"Zhou, Long, and Leonid A. Sazanov. “Structure and Conformational Plasticity of the Intact Thermus Thermophilus V/A-Type ATPase.” <i>Science</i>, vol. 365, no. 6455, eaaw9144, AAAS, 2019, doi:<a href=\"https://doi.org/10.1126/science.aaw9144\">10.1126/science.aaw9144</a>.","apa":"Zhou, L., &#38; Sazanov, L. A. (2019). Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase. <i>Science</i>. AAAS. <a href=\"https://doi.org/10.1126/science.aaw9144\">https://doi.org/10.1126/science.aaw9144</a>","ama":"Zhou L, Sazanov LA. Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase. <i>Science</i>. 2019;365(6455). doi:<a href=\"https://doi.org/10.1126/science.aaw9144\">10.1126/science.aaw9144</a>","short":"L. Zhou, L.A. Sazanov, Science 365 (2019).","ieee":"L. Zhou and L. A. Sazanov, “Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase,” <i>Science</i>, vol. 365, no. 6455. AAAS, 2019."},"issue":"6455","month":"08","article_number":"eaaw9144","department":[{"_id":"LeSa"}],"acknowledged_ssus":[{"_id":"ScienComp"}],"intvolume":"       365","abstract":[{"lang":"eng","text":"V (vacuolar)/A (archaeal)-type adenosine triphosphatases (ATPases), found in archaeaand eubacteria, couple ATP hydrolysis or synthesis to proton translocation across theplasma membrane using the rotary-catalysis mechanism. They belong to the V-typeATPase family, which differs from the mitochondrial/chloroplast F-type ATP synthasesin overall architecture. We solved cryo–electron microscopy structures of the intactThermus thermophilusV/A-ATPase, reconstituted into lipid nanodiscs, in three rotationalstates and two substates. These structures indicate substantial flexibility betweenV1and Voin a working enzyme, which results from mechanical competition between centralshaft rotation and resistance from the peripheral stalks. We also describedetails of adenosine diphosphate inhibition release, V1-Votorque transmission, andproton translocation, which are relevant for the entire V-type ATPase family."}],"publication_identifier":{"issn":["0036-8075"],"eissn":["1095-9203"]},"publication_status":"published","oa_version":"None","title":"Structure and conformational plasticity of the intact Thermus thermophilus V/A-type ATPase","scopus_import":"1","day":"23","author":[{"last_name":"Zhou","full_name":"Zhou, Long","id":"3E751364-F248-11E8-B48F-1D18A9856A87","first_name":"Long","orcid":"0000-0002-1864-8951"},{"full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989"}],"date_created":"2019-09-07T19:04:45Z","volume":365,"status":"public","publication":"Science","date_published":"2019-08-23T00:00:00Z","pmid":1,"related_material":{"link":[{"description":"News on IST Website","url":"https://ist.ac.at/en/news/structure-of-protein-nano-turbine-revealed/","relation":"press_release"}]},"external_id":{"pmid":["31439765"],"isi":["000482464000043"]},"isi":1,"year":"2019","quality_controlled":"1","publisher":"AAAS","article_processing_charge":"No","doi":"10.1126/science.aaw9144","type":"journal_article","_id":"6859","date_updated":"2023-08-29T07:52:02Z"}]