한빛사논문
- 조회929
Song-Yi Lee1,8, Joleen S. Cheah2,8, Boxuan Zhao1, Charles Xu3, Heegwang Roh 4, Christina K. Kim1,6, Kelvin F. Cho1,7, Namrata D. Udeshi 3, Steven A. Carr3 & Alice Y. Ting 1,2,4,5
1Department of Genetics, Stanford University, Stanford, CA, USA.
2Department of Biology, Stanford University, Stanford, CA, USA.
3Broad Institute of MIT and Harvard, Cambridge, MA, USA.
4Department of Chemistry, Stanford University, Stanford, CA, USA.
5Chan Zuckerberg Biohub–San Francisco, San Francisco, CA, USA.
6Present address: Center for Neuroscience and Department of Neurology, University of California, Davis, CA, USA.
7Present address: Amgen Research, South San Francisco, CA, USA.
8These authors contributed equally: Song-Yi Lee, Joleen S. Cheah.
Corresponding author : Correspondence to Alice Y. Ting.
Abstract
The incorporation of light-responsive domains into engineered proteins has enabled control of protein localization, interactions and function with light. We integrated optogenetic control into proximity labeling, a cornerstone technique for high-resolution proteomic mapping of organelles and interactomes in living cells. Through structure-guided screening and directed evolution, we installed the light-sensitive LOV domain into the proximity labeling enzyme TurboID to rapidly and reversibly control its labeling activity with low-power blue light. 'LOV-Turbo' works in multiple contexts and dramatically reduces background in biotin-rich environments such as neurons. We used LOV-Turbo for pulse-chase labeling to discover proteins that traffic between endoplasmic reticulum, nuclear and mitochondrial compartments under cellular stress. We also showed that instead of external light, LOV-Turbo can be activated by bioluminescence resonance energy transfer from luciferase, enabling interaction-dependent proximity labeling. Overall, LOV-Turbo increases the spatial and temporal precision of proximity labeling, expanding the scope of experimental questions that can be addressed with proximity labeling.
논문정보
- Research article
- 2023년 05월 (BRIC 등록일 2023-05-18)
- 국외 연구진
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