[{"title":"Electron microscopic detection of single membrane proteins by a specific chemical labeling","file":[{"creator":"dernst","file_size":7197776,"relation":"main_file","content_type":"application/pdf","file_name":"2019_iScience_Tabata.pdf","access_level":"open_access","date_created":"2020-02-04T10:48:36Z","checksum":"f3e90056a49f09b205b1c4f8c739ffd1","file_id":"7448","date_updated":"2020-07-14T12:47:57Z"}],"day":"20","author":[{"id":"4427179E-F248-11E8-B48F-1D18A9856A87","full_name":"Tabata, Shigekazu","last_name":"Tabata","first_name":"Shigekazu"},{"full_name":"Jevtic, Marijo","id":"4BE3BC94-F248-11E8-B48F-1D18A9856A87","last_name":"Jevtic","first_name":"Marijo"},{"first_name":"Nobutaka","last_name":"Kurashige","full_name":"Kurashige, Nobutaka"},{"last_name":"Fuchida","first_name":"Hirokazu","full_name":"Fuchida, Hirokazu"},{"first_name":"Munetsugu","last_name":"Kido","full_name":"Kido, Munetsugu"},{"last_name":"Tani","first_name":"Kazushi","full_name":"Tani, Kazushi"},{"full_name":"Zenmyo, Naoki","last_name":"Zenmyo","first_name":"Naoki"},{"last_name":"Uchinomiya","first_name":"Shohei","full_name":"Uchinomiya, Shohei"},{"orcid":"0000-0001-7429-7896","id":"2E55CDF2-F248-11E8-B48F-1D18A9856A87","full_name":"Harada, Harumi","first_name":"Harumi","last_name":"Harada"},{"full_name":"Itakura, Makoto","last_name":"Itakura","first_name":"Makoto"},{"full_name":"Hamachi, Itaru","last_name":"Hamachi","first_name":"Itaru"},{"last_name":"Shigemoto","first_name":"Ryuichi","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444","full_name":"Shigemoto, Ryuichi"},{"first_name":"Akio","last_name":"Ojida","full_name":"Ojida, Akio"}],"tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"},"article_type":"original","scopus_import":"1","ec_funded":1,"article_processing_charge":"No","publication":"iScience","department":[{"_id":"RySh"}],"pmid":1,"publisher":"Elsevier","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","quality_controlled":"1","doi":"10.1016/j.isci.2019.11.025","publication_identifier":{"issn":["2589-0042"]},"issue":"12","language":[{"iso":"eng"}],"project":[{"name":"In situ analysis of single channel subunit composition in neurons: physiological implication in synaptic plasticity and behaviour","_id":"25CA28EA-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","grant_number":"694539"},{"grant_number":"720270","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)","_id":"25CBA828-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"}],"date_created":"2020-01-29T15:56:56Z","file_date_updated":"2020-07-14T12:47:57Z","volume":22,"type":"journal_article","month":"12","oa_version":"Published Version","abstract":[{"lang":"eng","text":"Electron microscopy (EM) is a technology that enables visualization of single proteins at a nanometer resolution. However, current protein analysis by EM mainly relies on immunolabeling with gold-particle-conjugated antibodies, which is compromised by large size of antibody, precluding precise detection of protein location in biological samples. Here, we develop a specific chemical labeling method for EM detection of proteins at single-molecular level. Rational design of α-helical peptide tag and probe structure provided a complementary reaction pair that enabled specific cysteine conjugation of the tag. The developed chemical labeling with gold-nanoparticle-conjugated probe showed significantly higher labeling efficiency and detectability of high-density clusters of tag-fused G protein-coupled receptors in freeze-fracture replicas compared with immunogold labeling. Furthermore, in ultrathin sections, the spatial resolution of the chemical labeling was significantly higher than that of antibody-mediated labeling. These results demonstrate substantial advantages of the chemical labeling approach for single protein visualization by EM."}],"date_updated":"2024-03-25T23:30:07Z","page":"256-268","_id":"7391","year":"2019","ddc":["570"],"date_published":"2019-12-20T00:00:00Z","has_accepted_license":"1","oa":1,"publication_status":"published","citation":{"short":"S. Tabata, M. Jevtic, N. Kurashige, H. Fuchida, M. Kido, K. Tani, N. Zenmyo, S. Uchinomiya, H. Harada, M. Itakura, I. Hamachi, R. Shigemoto, A. Ojida, IScience 22 (2019) 256–268.","chicago":"Tabata, Shigekazu, Marijo Jevtic, Nobutaka Kurashige, Hirokazu Fuchida, Munetsugu Kido, Kazushi Tani, Naoki Zenmyo, et al. “Electron Microscopic Detection of Single Membrane Proteins by a Specific Chemical Labeling.” <i>IScience</i>. Elsevier, 2019. <a href=\"https://doi.org/10.1016/j.isci.2019.11.025\">https://doi.org/10.1016/j.isci.2019.11.025</a>.","ieee":"S. Tabata <i>et al.</i>, “Electron microscopic detection of single membrane proteins by a specific chemical labeling,” <i>iScience</i>, vol. 22, no. 12. Elsevier, pp. 256–268, 2019.","apa":"Tabata, S., Jevtic, M., Kurashige, N., Fuchida, H., Kido, M., Tani, K., … Ojida, A. (2019). Electron microscopic detection of single membrane proteins by a specific chemical labeling. <i>IScience</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.isci.2019.11.025\">https://doi.org/10.1016/j.isci.2019.11.025</a>","mla":"Tabata, Shigekazu, et al. “Electron Microscopic Detection of Single Membrane Proteins by a Specific Chemical Labeling.” <i>IScience</i>, vol. 22, no. 12, Elsevier, 2019, pp. 256–68, doi:<a href=\"https://doi.org/10.1016/j.isci.2019.11.025\">10.1016/j.isci.2019.11.025</a>.","ista":"Tabata S, Jevtic M, Kurashige N, Fuchida H, Kido M, Tani K, Zenmyo N, Uchinomiya S, Harada H, Itakura M, Hamachi I, Shigemoto R, Ojida A. 2019. Electron microscopic detection of single membrane proteins by a specific chemical labeling. iScience. 22(12), 256–268.","ama":"Tabata S, Jevtic M, Kurashige N, et al. Electron microscopic detection of single membrane proteins by a specific chemical labeling. <i>iScience</i>. 2019;22(12):256-268. doi:<a href=\"https://doi.org/10.1016/j.isci.2019.11.025\">10.1016/j.isci.2019.11.025</a>"},"related_material":{"record":[{"status":"public","id":"11393","relation":"dissertation_contains"}]},"intvolume":"        22","external_id":{"isi":[":000504652000020"],"pmid":["31786521"]},"status":"public"},{"department":[{"_id":"SiHi"}],"publisher":"Elsevier","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","article_processing_charge":"No","tmp":{"name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)","short":"CC BY-NC-ND (4.0)","image":"/images/cc_by_nc_nd.png","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode"},"publication":"iScience","day":"31","file":[{"file_name":"2019_iScience_Amberg.pdf","creator":"dernst","file_size":8365970,"content_type":"application/pdf","relation":"main_file","checksum":"a9ad2296726c9474ad5860c9c2f53622","file_id":"6452","date_updated":"2020-07-14T12:47:30Z","access_level":"open_access","date_created":"2019-05-14T11:51:51Z"}],"author":[{"first_name":"Nicole","last_name":"Amberg","id":"4CD6AAC6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-3183-8207","full_name":"Amberg, Nicole"},{"first_name":"Panagiota A.","last_name":"Sotiropoulou","full_name":"Sotiropoulou, Panagiota A."},{"last_name":"Heller","first_name":"Gerwin","full_name":"Heller, Gerwin"},{"full_name":"Lichtenberger, Beate M.","first_name":"Beate M.","last_name":"Lichtenberger"},{"last_name":"Holcmann","first_name":"Martin","full_name":"Holcmann, Martin"},{"full_name":"Camurdanoglu, Bahar","first_name":"Bahar","last_name":"Camurdanoglu"},{"first_name":"Temenuschka","last_name":"Baykuscheva-Gentscheva","full_name":"Baykuscheva-Gentscheva, Temenuschka"},{"first_name":"Cedric","last_name":"Blanpain","full_name":"Blanpain, Cedric"},{"full_name":"Sibilia, Maria","last_name":"Sibilia","first_name":"Maria"}],"title":"EGFR controls hair shaft differentiation in a p53-independent manner","language":[{"iso":"eng"}],"isi":1,"publication_identifier":{"issn":["2589-0042"]},"quality_controlled":"1","doi":"10.1016/j.isci.2019.04.018","year":"2019","_id":"6451","date_updated":"2023-09-08T11:38:04Z","abstract":[{"text":"Epidermal growth factor receptor (EGFR) signaling controls skin development and homeostasis inmice and humans, and its deficiency causes severe skin inflammation, which might affect epidermalstem cell behavior. Here, we describe the inflammation-independent effects of EGFR deficiency dur-ing skin morphogenesis and in adult hair follicle stem cells. Expression and alternative splicing analysisof RNA sequencing data from interfollicular epidermis and outer root sheath indicate that EGFR con-trols genes involved in epidermal differentiation and also in centrosome function, DNA damage, cellcycle, and apoptosis. Genetic experiments employingp53deletion in EGFR-deficient epidermis revealthat EGFR signaling exhibitsp53-dependent functions in proliferative epidermal compartments, aswell asp53-independent functions in differentiated hair shaft keratinocytes. Loss of EGFR leads toabsence of LEF1 protein specifically in the innermost epithelial hair layers, resulting in disorganizationof medulla cells. Thus, our results uncover important spatial and temporal features of cell-autonomousEGFR functions in the epidermis.","lang":"eng"}],"type":"journal_article","month":"05","oa_version":"Published Version","page":"243-256","date_created":"2019-05-14T11:47:40Z","file_date_updated":"2020-07-14T12:47:30Z","volume":15,"status":"public","external_id":{"isi":["000470104600022"]},"citation":{"short":"N. Amberg, P.A. Sotiropoulou, G. Heller, B.M. Lichtenberger, M. Holcmann, B. Camurdanoglu, T. Baykuscheva-Gentscheva, C. Blanpain, M. Sibilia, IScience 15 (2019) 243–256.","ieee":"N. Amberg <i>et al.</i>, “EGFR controls hair shaft differentiation in a p53-independent manner,” <i>iScience</i>, vol. 15. Elsevier, pp. 243–256, 2019.","chicago":"Amberg, Nicole, Panagiota A. Sotiropoulou, Gerwin Heller, Beate M. Lichtenberger, Martin Holcmann, Bahar Camurdanoglu, Temenuschka Baykuscheva-Gentscheva, Cedric Blanpain, and Maria Sibilia. “EGFR Controls Hair Shaft Differentiation in a P53-Independent Manner.” <i>IScience</i>. Elsevier, 2019. <a href=\"https://doi.org/10.1016/j.isci.2019.04.018\">https://doi.org/10.1016/j.isci.2019.04.018</a>.","mla":"Amberg, Nicole, et al. “EGFR Controls Hair Shaft Differentiation in a P53-Independent Manner.” <i>IScience</i>, vol. 15, Elsevier, 2019, pp. 243–56, doi:<a href=\"https://doi.org/10.1016/j.isci.2019.04.018\">10.1016/j.isci.2019.04.018</a>.","ista":"Amberg N, Sotiropoulou PA, Heller G, Lichtenberger BM, Holcmann M, Camurdanoglu B, Baykuscheva-Gentscheva T, Blanpain C, Sibilia M. 2019. EGFR controls hair shaft differentiation in a p53-independent manner. iScience. 15, 243–256.","apa":"Amberg, N., Sotiropoulou, P. A., Heller, G., Lichtenberger, B. M., Holcmann, M., Camurdanoglu, B., … Sibilia, M. (2019). EGFR controls hair shaft differentiation in a p53-independent manner. <i>IScience</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.isci.2019.04.018\">https://doi.org/10.1016/j.isci.2019.04.018</a>","ama":"Amberg N, Sotiropoulou PA, Heller G, et al. EGFR controls hair shaft differentiation in a p53-independent manner. <i>iScience</i>. 2019;15:243-256. doi:<a href=\"https://doi.org/10.1016/j.isci.2019.04.018\">10.1016/j.isci.2019.04.018</a>"},"intvolume":"        15","has_accepted_license":"1","publication_status":"published","oa":1,"date_published":"2019-05-31T00:00:00Z","ddc":["570"]}]