---
_id: '6564'
abstract:
- lang: eng
  text: Optogenetics enables the spatio-temporally precise control of cell and animal
    behavior. Many optogenetic tools are driven by light-controlled protein–protein
    interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs).
    Applying light-controlled PPIs to new target proteins is challenging because it
    is difficult to predict which of the many available LSDs, if any, will yield robust
    light regulation. As a consequence, fusion protein libraries need to be prepared
    and tested, but methods and platforms to facilitate this process are currently
    not available. Here, we developed a genetic engineering strategy and vector library
    for the rapid generation of light-controlled PPIs. The strategy permits fusing
    a target protein to multiple LSDs efficiently and in two orientations. The public
    and expandable library contains 29 vectors with blue, green or red light-responsive
    LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate
    the versatility of the approach and the necessity for sampling LSDs by generating
    light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a
    new resource for optical regulation of a broad range of target proteins in cell
    and developmental biology.
article_processing_charge: No
article_type: original
author:
- first_name: Alexandra-Madelaine
  full_name: Tichy, Alexandra-Madelaine
  id: 29D8BB2C-F248-11E8-B48F-1D18A9856A87
  last_name: Tichy
- first_name: Elliot J.
  full_name: Gerrard, Elliot J.
  last_name: Gerrard
- first_name: Julien M.D.
  full_name: Legrand, Julien M.D.
  last_name: Legrand
- first_name: Robin M.
  full_name: Hobbs, Robin M.
  last_name: Hobbs
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. Engineering strategy
    and vector library for the rapid generation of modular light-controlled protein–protein
    interactions. <i>Journal of Molecular Biology</i>. 2019;431(17):3046-3055. doi:<a
    href="https://doi.org/10.1016/j.jmb.2019.05.033">10.1016/j.jmb.2019.05.033</a>
  apa: Tichy, A.-M., Gerrard, E. J., Legrand, J. M. D., Hobbs, R. M., &#38; Janovjak,
    H. L. (2019). Engineering strategy and vector library for the rapid generation
    of modular light-controlled protein–protein interactions. <i>Journal of Molecular
    Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jmb.2019.05.033">https://doi.org/10.1016/j.jmb.2019.05.033</a>
  chicago: Tichy, Alexandra-Madelaine, Elliot J. Gerrard, Julien M.D. Legrand, Robin
    M. Hobbs, and Harald L Janovjak. “Engineering Strategy and Vector Library for
    the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.”
    <i>Journal of Molecular Biology</i>. Elsevier, 2019. <a href="https://doi.org/10.1016/j.jmb.2019.05.033">https://doi.org/10.1016/j.jmb.2019.05.033</a>.
  ieee: A.-M. Tichy, E. J. Gerrard, J. M. D. Legrand, R. M. Hobbs, and H. L. Janovjak,
    “Engineering strategy and vector library for the rapid generation of modular light-controlled
    protein–protein interactions,” <i>Journal of Molecular Biology</i>, vol. 431,
    no. 17. Elsevier, pp. 3046–3055, 2019.
  ista: Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. 2019. Engineering
    strategy and vector library for the rapid generation of modular light-controlled
    protein–protein interactions. Journal of Molecular Biology. 431(17), 3046–3055.
  mla: Tichy, Alexandra-Madelaine, et al. “Engineering Strategy and Vector Library
    for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.”
    <i>Journal of Molecular Biology</i>, vol. 431, no. 17, Elsevier, 2019, pp. 3046–55,
    doi:<a href="https://doi.org/10.1016/j.jmb.2019.05.033">10.1016/j.jmb.2019.05.033</a>.
  short: A.-M. Tichy, E.J. Gerrard, J.M.D. Legrand, R.M. Hobbs, H.L. Janovjak, Journal
    of Molecular Biology 431 (2019) 3046–3055.
date_created: 2019-06-16T21:59:14Z
date_published: 2019-08-09T00:00:00Z
date_updated: 2023-08-28T09:39:22Z
day: '09'
department:
- _id: HaJa
doi: 10.1016/j.jmb.2019.05.033
external_id:
  isi:
  - '000482872100002'
intvolume: '       431'
isi: 1
issue: '17'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: http://www.biorxiv.org/content/10.1101/583369v1
month: '08'
oa: 1
oa_version: Preprint
page: 3046-3055
publication: Journal of Molecular Biology
publication_identifier:
  eissn:
  - '10898638'
  issn:
  - '00222836'
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Engineering strategy and vector library for the rapid generation of modular
  light-controlled protein–protein interactions
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 431
year: '2019'
...
---
_id: '5984'
abstract:
- lang: eng
  text: G-protein-coupled receptors (GPCRs) form the largest receptor family, relay
    environmental stimuli to changes in cell behavior and represent prime drug targets.
    Many GPCRs are classified as orphan receptors because of the limited knowledge
    on their ligands and coupling to cellular signaling machineries. Here, we engineer
    a library of 63 chimeric receptors that contain the signaling domains of human
    orphan and understudied GPCRs functionally linked to the light-sensing domain
    of rhodopsin. Upon stimulation with visible light, we identify activation of canonical
    cell signaling pathways, including cAMP-, Ca2+-, MAPK/ERK-, and Rho-dependent
    pathways, downstream of the engineered receptors. For the human pseudogene GPR33,
    we resurrect a signaling function that supports its hypothesized role as a pathogen
    entry site. These results demonstrate that substituting unknown chemical activators
    with a light switch can reveal information about protein function and provide
    an optically controlled protein library for exploring the physiology and therapeutic
    potential of understudied GPCRs.
article_number: '1950'
article_processing_charge: No
author:
- first_name: Maurizio
  full_name: Morri, Maurizio
  id: 4863116E-F248-11E8-B48F-1D18A9856A87
  last_name: Morri
- first_name: Inmaculada
  full_name: Sanchez-Romero, Inmaculada
  id: 3D9C5D30-F248-11E8-B48F-1D18A9856A87
  last_name: Sanchez-Romero
- first_name: Alexandra-Madelaine
  full_name: Tichy, Alexandra-Madelaine
  id: 29D8BB2C-F248-11E8-B48F-1D18A9856A87
  last_name: Tichy
- first_name: Stephanie
  full_name: Kainrath, Stephanie
  id: 32CFBA64-F248-11E8-B48F-1D18A9856A87
  last_name: Kainrath
- first_name: Elliot J.
  full_name: Gerrard, Elliot J.
  last_name: Gerrard
- first_name: Priscila
  full_name: Hirschfeld, Priscila
  id: 435ACB3A-F248-11E8-B48F-1D18A9856A87
  last_name: Hirschfeld
- first_name: Jan
  full_name: Schwarz, Jan
  id: 346C1EC6-F248-11E8-B48F-1D18A9856A87
  last_name: Schwarz
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Morri M, Sanchez-Romero I, Tichy A-M, et al. Optical functionalization of human
    class A orphan G-protein-coupled receptors. <i>Nature Communications</i>. 2018;9(1).
    doi:<a href="https://doi.org/10.1038/s41467-018-04342-1">10.1038/s41467-018-04342-1</a>
  apa: Morri, M., Sanchez-Romero, I., Tichy, A.-M., Kainrath, S., Gerrard, E. J.,
    Hirschfeld, P., … Janovjak, H. L. (2018). Optical functionalization of human class
    A orphan G-protein-coupled receptors. <i>Nature Communications</i>. Springer Nature.
    <a href="https://doi.org/10.1038/s41467-018-04342-1">https://doi.org/10.1038/s41467-018-04342-1</a>
  chicago: Morri, Maurizio, Inmaculada Sanchez-Romero, Alexandra-Madelaine Tichy,
    Stephanie Kainrath, Elliot J. Gerrard, Priscila Hirschfeld, Jan Schwarz, and Harald
    L Janovjak. “Optical Functionalization of Human Class A Orphan G-Protein-Coupled
    Receptors.” <i>Nature Communications</i>. Springer Nature, 2018. <a href="https://doi.org/10.1038/s41467-018-04342-1">https://doi.org/10.1038/s41467-018-04342-1</a>.
  ieee: M. Morri <i>et al.</i>, “Optical functionalization of human class A orphan
    G-protein-coupled receptors,” <i>Nature Communications</i>, vol. 9, no. 1. Springer
    Nature, 2018.
  ista: Morri M, Sanchez-Romero I, Tichy A-M, Kainrath S, Gerrard EJ, Hirschfeld P,
    Schwarz J, Janovjak HL. 2018. Optical functionalization of human class A orphan
    G-protein-coupled receptors. Nature Communications. 9(1), 1950.
  mla: Morri, Maurizio, et al. “Optical Functionalization of Human Class A Orphan
    G-Protein-Coupled Receptors.” <i>Nature Communications</i>, vol. 9, no. 1, 1950,
    Springer Nature, 2018, doi:<a href="https://doi.org/10.1038/s41467-018-04342-1">10.1038/s41467-018-04342-1</a>.
  short: M. Morri, I. Sanchez-Romero, A.-M. Tichy, S. Kainrath, E.J. Gerrard, P. Hirschfeld,
    J. Schwarz, H.L. Janovjak, Nature Communications 9 (2018).
date_created: 2019-02-14T10:50:24Z
date_published: 2018-12-01T00:00:00Z
date_updated: 2023-09-19T14:29:32Z
day: '01'
ddc:
- '570'
department:
- _id: HaJa
- _id: CaGu
- _id: MiSi
doi: 10.1038/s41467-018-04342-1
ec_funded: 1
external_id:
  isi:
  - '000432280000006'
file:
- access_level: open_access
  checksum: 8325fcc194264af4749e662a73bf66b5
  content_type: application/pdf
  creator: kschuh
  date_created: 2019-02-14T10:58:29Z
  date_updated: 2020-07-14T12:47:14Z
  file_id: '5985'
  file_name: 2018_Springer_Morri.pdf
  file_size: 1349914
  relation: main_file
file_date_updated: 2020-07-14T12:47:14Z
has_accepted_license: '1'
intvolume: '         9'
isi: 1
issue: '1'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
project:
- _id: 25548C20-B435-11E9-9278-68D0E5697425
  call_identifier: FP7
  grant_number: '303564'
  name: Microbial Ion Channels for Synthetic Neurobiology
- _id: 255A6082-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: W1232-B24
  name: Molecular Drug Targets
publication: Nature Communications
publication_identifier:
  issn:
  - 2041-1723
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Optical functionalization of human class A orphan G-protein-coupled receptors
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 9
year: '2018'
...
---
_id: '1441'
abstract:
- lang: eng
  text: 'Optogenetics and photopharmacology enable the spatio-temporal control of
    cell and animal behavior by light. Although red light offers deep-tissue penetration
    and minimal phototoxicity, very few red-light-sensitive optogenetic methods are
    currently available. We have now developed a red-light-induced homodimerization
    domain. We first showed that an optimized sensory domain of the cyanobacterial
    phytochrome 1 can be expressed robustly and without cytotoxicity in human cells.
    We then applied this domain to induce the dimerization of two receptor tyrosine
    kinases—the fibroblast growth factor receptor 1 and the neurotrophin receptor
    trkB. This new optogenetic method was then used to activate the MAPK/ERK pathway
    non-invasively in mammalian tissue and in multicolor cell-signaling experiments.
    The light-controlled dimerizer and red-light-activated receptor tyrosine kinases
    will prove useful to regulate a variety of cellular processes with light. Go deep
    with red: The sensory domain (S) of the cyanobacterial phytochrome 1 (CPH1) was
    repurposed to induce the homodimerization of proteins in living cells by red light.
    By using this domain, light-activated protein kinases were engineered that can
    be activated orthogonally from many fluorescent proteins and through mammalian
    tissue. Pr/Pfr=red-/far-red-absorbing state of CPH1.'
acknowledgement: 'A.I.-P. was supported by a Ramon Areces fellowship, and E.R. by
  the graduate program MolecularDrugTargets (Austrian Science Fund (FWF): W1232) and
  a FemTech fellowship (Austrian Research Promotion Agency: 3580812).'
author:
- first_name: Eva
  full_name: Gschaider-Reichhart, Eva
  id: 3FEE232A-F248-11E8-B48F-1D18A9856A87
  last_name: Gschaider-Reichhart
  orcid: 0000-0002-7218-7738
- first_name: Álvaro
  full_name: Inglés Prieto, Álvaro
  id: 2A9DB292-F248-11E8-B48F-1D18A9856A87
  last_name: Inglés Prieto
  orcid: 0000-0002-5409-8571
- first_name: Alexandra-Madelaine
  full_name: Tichy, Alexandra-Madelaine
  id: 29D8BB2C-F248-11E8-B48F-1D18A9856A87
  last_name: Tichy
- first_name: Catherine
  full_name: Mckenzie, Catherine
  id: 3EEDE19A-F248-11E8-B48F-1D18A9856A87
  last_name: Mckenzie
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Gschaider-Reichhart E, Inglés Prieto Á, Tichy A-M, Mckenzie C, Janovjak HL.
    A phytochrome sensory domain permits receptor activation by red light. <i>Angewandte
    Chemie - International Edition</i>. 2016;55(21):6339-6342. doi:<a href="https://doi.org/10.1002/anie.201601736">10.1002/anie.201601736</a>
  apa: Gschaider-Reichhart, E., Inglés Prieto, Á., Tichy, A.-M., Mckenzie, C., &#38;
    Janovjak, H. L. (2016). A phytochrome sensory domain permits receptor activation
    by red light. <i>Angewandte Chemie - International Edition</i>. Wiley. <a href="https://doi.org/10.1002/anie.201601736">https://doi.org/10.1002/anie.201601736</a>
  chicago: Gschaider-Reichhart, Eva, Álvaro Inglés Prieto, Alexandra-Madelaine Tichy,
    Catherine Mckenzie, and Harald L Janovjak. “A Phytochrome Sensory Domain Permits
    Receptor Activation by Red Light.” <i>Angewandte Chemie - International Edition</i>.
    Wiley, 2016. <a href="https://doi.org/10.1002/anie.201601736">https://doi.org/10.1002/anie.201601736</a>.
  ieee: E. Gschaider-Reichhart, Á. Inglés Prieto, A.-M. Tichy, C. Mckenzie, and H.
    L. Janovjak, “A phytochrome sensory domain permits receptor activation by red
    light,” <i>Angewandte Chemie - International Edition</i>, vol. 55, no. 21. Wiley,
    pp. 6339–6342, 2016.
  ista: Gschaider-Reichhart E, Inglés Prieto Á, Tichy A-M, Mckenzie C, Janovjak HL.
    2016. A phytochrome sensory domain permits receptor activation by red light. Angewandte
    Chemie - International Edition. 55(21), 6339–6342.
  mla: Gschaider-Reichhart, Eva, et al. “A Phytochrome Sensory Domain Permits Receptor
    Activation by Red Light.” <i>Angewandte Chemie - International Edition</i>, vol.
    55, no. 21, Wiley, 2016, pp. 6339–42, doi:<a href="https://doi.org/10.1002/anie.201601736">10.1002/anie.201601736</a>.
  short: E. Gschaider-Reichhart, Á. Inglés Prieto, A.-M. Tichy, C. Mckenzie, H.L.
    Janovjak, Angewandte Chemie - International Edition 55 (2016) 6339–6342.
date_created: 2018-12-11T11:52:02Z
date_published: 2016-05-17T00:00:00Z
date_updated: 2023-09-07T12:49:08Z
day: '17'
ddc:
- '571'
- '576'
department:
- _id: HaJa
doi: 10.1002/anie.201601736
ec_funded: 1
file:
- access_level: open_access
  checksum: 26da07960e57ac4750b54179197ce57f
  content_type: application/pdf
  creator: system
  date_created: 2018-12-12T10:17:03Z
  date_updated: 2020-07-14T12:44:55Z
  file_id: '5255'
  file_name: IST-2017-840-v1+1_reichhart.pdf
  file_size: 1268662
  relation: main_file
file_date_updated: 2020-07-14T12:44:55Z
has_accepted_license: '1'
intvolume: '        55'
issue: '21'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Submitted Version
page: 6339 - 6342
project:
- _id: 25548C20-B435-11E9-9278-68D0E5697425
  call_identifier: FP7
  grant_number: '303564'
  name: Microbial Ion Channels for Synthetic Neurobiology
- _id: 255A6082-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: W1232-B24
  name: Molecular Drug Targets
publication: Angewandte Chemie - International Edition
publication_status: published
publisher: Wiley
publist_id: '5755'
pubrep_id: '840'
quality_controlled: '1'
related_material:
  record:
  - id: '418'
    relation: dissertation_contains
    status: public
scopus_import: 1
status: public
title: A phytochrome sensory domain permits receptor activation by red light
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 55
year: '2016'
...