Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity

• Type of Material: Article
• Authors: Mark Shaaya, University of Illinois; Jordan Fauser, University of Illinois; Anastasia Zhurikhina, Georgia Institute of Technology; Jason E. Conage-Pough, Massachusetts Institute of Technology; Vincent Huyot, University of Illinois; Martin Brennan, University of Illinois; Cameron T. Flower, Massachusetts Institute of Technology; Jacob Matsche, University of Illinois; Shahzeb Khan, University of Illinois; Viswanathan Natarajan, University of Illinois; Jalees Rehman, University of Illinois; Pradeep Kota, University of North Carolina; Forest M. White, Massachusetts Institute of Technology; Denis Tsygankov, Emory University; Andrei V. Karginov, University of Illinois
• Language: English
• Date: 2020-09-23
• Publisher: ELIFE SCIENCES PUBLICATIONS LTD
• Publication Version: Final Published Version
• Copyright Statement: © 2020, Shaaya et al
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.7554/eLife.60647
• Title of Journal or Parent Work: ELIFE
• Volume: 9
• Start Page: 1
• End Page: 73
• Grant/Funding Information: We are grateful to the NIH and Chicago Biomedical Consortium for the funding (CBC Pilot Grant, R21CA212907, R21CA159179, and R01GM118582 grants to AVK; R21CA223915 to AVK and JR; training grant T32 HL007829-22 to MS, JF, and MB; and P01 HL060678 to AVK, VN and JR). We are also thankful to NCI for the funding (CA210180 and CA238720 grants to FMW) and for the U.S. Army Research Office (ARO) (W911NF-17-1-0395 to DT).
• Keywords: RECEPTOR TYROSINE KINASES; OPTOGENETIC CONTROL; GENE-EXPRESSION; Life Sciences & Biomedicine - Other Topics; SIGNALING DYNAMICS; C-SRC; PROTEIN-KINASES; Biology; Life Sciences & Biomedicine; Science & Technology; COMPUTATIONAL PLATFORM; SITE-SPECIFIC RECOMBINATION; CELL-MIGRATION; SPATIOTEMPORAL CONTROL
• Research Categories: Biology, Genetics; Biology, Cell

Abstract

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.