Optogenetic activation of heterotrimeric G-proteins by LOV2GIVe, a rationally engineered modular protein

• Published in: eLife Vol. 9
• Main Authors: Garcia-Marcos, Mikel, Parag-Sharma, Kshitij, Marivin, Arthur, Maziarz, Marcin, Luebbers, Alex, Nguyen, Lien T
• Format: Journal Article
• Language: English
• Published: England eLife Science Publications, Ltd 16.09.2020; eLife Sciences Publications, Ltd; eLife Sciences Publications Ltd
• Subjects: Animals; Avena - genetics; Biochemistry and Chemical Biology; Cell Biology; Escherichia coli - genetics; GEF; Girdin; GIV; GPCR; GTPase; Health aspects; Heterotrimeric GTP-Binding Proteins - chemistry; Heterotrimeric GTP-Binding Proteins - genetics; Heterotrimeric GTP-Binding Proteins - metabolism; Humans; Optogenetics; Optogenetics - methods; Plant Proteins - chemistry; Plant Proteins - genetics; Plant Proteins - metabolism; Protein Engineering - methods; Proteins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Saccharomyces cerevisiae - genetics; Tools and Resources; cell biology; chemical biology; GIV; biochemistry; GEF; GPCR; Optogenetics; Girdin; GTPase; human; E. coli; S. cerevisiae
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.60155

Heterotrimeric G-proteins are signal transducers involved in mediating the action of many natural extracellular stimuli and 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, that is metazoan opsins, which are light-activated G-protein-coupled receptors (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.