|Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity.
|Year of Publication
|Shaaya M, Fauser J, Zhurikhina A, Conage-Pough JE, Huyot V, Brennan M, Flower CT, Matsche J, Khan S, Natarajan V, Rehman J, Kota P, White FM, Tsygankov D, Karginov AV
|2020 09 23
Engineered allosteric regulation of protein activity provides significant advantages for the development of robust and broadly applicable tools. However, the application of allosteric switches in optogenetics has been scarce and suffers from critical limitations. Here, we report an optogenetic approach that utilizes an engineered Light-Regulated (LightR) allosteric switch module to achieve tight spatiotemporal control of enzymatic activity. Using the tyrosine kinase Src as a model, we demonstrate efficient regulation of the kinase and identify temporally distinct signaling responses ranging from seconds to minutes. LightR-Src off-kinetics can be tuned by modulating the LightR photoconversion cycle. A fast cycling variant enables the stimulation of transient pulses and local regulation of activity in a selected region of a cell. The design of the LightR module ensures broad applicability of the tool, as we demonstrate by achieving light-mediated regulation of Abl and bRaf kinases as well as Cre recombinase.
|PubMed Central ID
|R21 CA212907 / CA / NCI NIH HHS / United States
T32 GM087237 / GM / NIGMS NIH HHS / United States
T32 HL007829 / HL / NHLBI NIH HHS / United States
W911NF-17-1-0395 / / Army Research Office / International
R21 CA159179 / CA / NCI NIH HHS / United States
P01 HL060678 / HL / NHLBI NIH HHS / United States
R21 CA223915 / CA / NCI NIH HHS / United States
U01 CA238720 / CA / NCI NIH HHS / United States
R01 GM118582 / GM / NIGMS NIH HHS / United States
U54 CA210180 / CA / NCI NIH HHS / United States