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

Mark Shaaya | Jordan Fauser | Anastasia Zhurikhina | Jason E Conage-Pough | Vincent Huyot | Martin Brennan | Cameron T Flower | Jacob Matsche | Shahzeb Khan | Viswanathan Natarajan | Jalees Rehman | Pradeep Kota | Forest M White | Denis Tsygankov | Andrei V Karginov
eLife | 2020

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 ID: 32965214

Associated grants

  • Agency: NCI NIH HHS, United States
    Id: R21 CA212907
  • Agency: NIGMS NIH HHS, United States
    Id: T32 GM087237
  • Agency: NHLBI NIH HHS, United States
    Id: T32 HL007829
  • Agency: Army Research Office, International
    Id: W911NF-17-1-0395
  • Agency: NCI NIH HHS, United States
    Id: R21 CA159179
  • Agency: NHLBI NIH HHS, United States
    Id: P01 HL060678
  • Agency: NCI NIH HHS, United States
    Id: R21 CA223915
  • Agency: NCI NIH HHS, United States
    Id: U01 CA238720
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM118582
  • Agency: NCI NIH HHS, United States
    Id: U54 CA210180

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.

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