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

Published: Aug. 19, 2020, 7:01 p.m.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.17.253781v1?rss=1 Authors: Garcia-Marcos, M., Parag-Sharma, K., Marivin, A., Maziarz, M., Luebbers, A., Nguyen, L. T. Abstract: Heterotrimeric G-proteins are signal transducers that mediate 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 engineered LOV2GIVe, a modular protein that allows the activation of Gi 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. To make LOV2GIVe, we fused a peptide derived from a non-GPCR protein that activates Gi (but not Gs, Gq, or G12) 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. Copy rights belong to original authors. Visit the link for more info