1. Biochemistry and Chemical Biology
  2. Cell Biology
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Optogenetic activation of heterotrimeric G-proteins by LOV2GIVe, a rationally engineered modular protein

  1. Mikel Garcia-Marcos  Is a corresponding author
  2. Kshitij Parag-Sharma
  3. Arthur Marivin
  4. Marcin Maziarz
  5. Alex Luebbers
  6. Lien T Nguyen
  1. Boston University School of Medicine, United States
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Cite this article as: eLife 2020;9:e60155 doi: 10.7554/eLife.60155

Abstract

Heterotrimeric G-proteins are signal transducers involved in mediating the action of many natural extracellular stimuli as well as of many therapeutic agents. Non-invasive approaches to manipulate the activity of G-proteins with high precision are crucial to understand their regulation in space and time. Here, we developed LOV2GIVe, an engineered modular protein that allows the activation of heterotrimeric G-proteins with blue light. This optogenetic construct relies on a versatile design that differs from tools previously developed for similar purposes, i.e. metazoan opsins, which are light-activated GPCRs. Instead, LOV2GIVe consists of the fusion of a G-protein activating peptide derived from a non-GPCR regulator of G-proteins to a small plant protein domain, such that light uncages the G-protein activating module. Targeting LOV2GIVe to cell membranes allowed for light-dependent activation of Gi proteins in different experimental systems. In summary, LOV2GIVe expands the armamentarium and versatility of tools available to manipulate heterotrimeric G-protein activity.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 2, 3 and 4 (and their corresponding supplements).

Article and author information

Author details

  1. Mikel Garcia-Marcos

    Department of Biochemistry, Boston University School of Medicine, Boston, United States
    For correspondence
    mgm1@bu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9513-4826
  2. Kshitij Parag-Sharma

    Department of Biochemistry, Boston University School of Medicine, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3638-0941
  3. Arthur Marivin

    Department of Biochemistry, Boston University School of Medicine, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Marcin Maziarz

    Department of Biochemistry, Boston University School of Medicine, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Alex Luebbers

    Department of Biochemistry, Boston University School of Medicine, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7733-4250
  6. Lien T Nguyen

    Biochemistry, Boston University School of Medicine, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.

Funding

National Institute of General Medical Sciences (R01GM136132)

  • Mikel Garcia-Marcos

American Cancer Society (PF-19-084-01-CDD)

  • Marcin Maziarz

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Volker Dötsch, Goethe University, Germany

Publication history

  1. Received: June 18, 2020
  2. Accepted: September 15, 2020
  3. Accepted Manuscript published: September 16, 2020 (version 1)
  4. Version of Record published: September 24, 2020 (version 2)

Copyright

© 2020, Garcia-Marcos et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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