Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity
Name
elife-60647-v4.pdf
Description
Published version
Size
5.61 MB
Format
Adobe PDF
Checksum (MD5)
be01be55c9e427466233d22583fa9659
Author(s)
Shaaya, Mark
Fauser, Jordan
Zhurikhina, Anastasia
Conage-Pough, Jason E
Huyot, Vincent
Brennan, Martin
Flower, Cameron T
Matsche, Jacob
Khan, Shahzeb
Natarajan, Viswanathan
Date Issued
2020
Journal
eLife
Publisher
eLife Sciences Publications, Ltd
Version
Final published version
Abstract
© 2020, eLife Sciences Publications Ltd. All rights reserved. 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.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
Center for Precision Cancer Medicine
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Computational and Systems Biology Program
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.7554/ELIFE.60647
