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

Elife. 2020; 
Mikel Garcia-Marcos, Kshitij Parag-Sharma, Arthur Marivin, Marcin Maziarz, Alex Luebbers, Lien T Nguyen
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GenParts™ DNA Fragments … Plasmid constructs The parental LOV2GIV sequence was obtained as a synthetic gene fragment from GenScript and subsequently amplified by PCR with extensions at … Purified His-Gai3 (100 nM) was diluted in assay buffer (20 mM Na- HEPES, pH 8, 100 mM NaCl, 1 mM EDTA … Get A Quote

摘要

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 derive... More

关键词

E. coli, GEF, GIV, GPCR, GTPase, Girdin, Optogenetics, S. cerevisiae, biochemistry, cell biology, chemical biology, human