Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Sep 23, 2020
- Source ID
- 10.7554/elife.60647
Entities
People
- Anastasia Zhurikhina
- Andrei V Karginov
- Cameron T Flower
- Denis Tsygankov
- Forest M. White
- Jacob Matsche
- Jalees Rehman
- Jason E Conage-Pough
- Jordan Fauser
- Mark Shaaya
- Martin Brennan
- Pradeep Kota
- Shahzeb Khan
- Vincent Huyot
- Viswanathan Natarajan
Organizations
- Army Research Office
- Georgia Tech
- Massachusetts Institute of Technology
- National Institutes of Health
- University of Illinois Urbana–Champaign
- University of Illinois at Chicago
- University of North Carolina