@article{651a5cad2ecb4374a5c0e233251d29a5,
title = "A genetically encoded photoproximity labeling approach for mapping protein territories",
abstract = "Studying dynamic biological processes requires approaches compatible with the lifetimes of the biochemical transactions under investigation, which can be very short. We describe a genetically encoded system that allows protein neighborhoods to be mapped using visible light. Our approach involves fusing an engineered flavoprotein to a protein of interest. Brief excitation of the fusion protein leads to the labeling of nearby proteins with cell-permeable probes. Mechanistic studies reveal different labeling pathways are operational depending on the nature of the exogenous probe that is employed. When combined with quantitative proteomics, this photoproximity labeling system generates “snapshots” of protein territories with high temporal and spatial resolution. The intrinsic fluorescence of the fusion domain permits correlated imaging and proteomics analyses, a capability that is exploited in several contexts, including defining the protein clients of the major vault protein. The technology should be broadly useful in the biomedical area.",
keywords = "LOV domain, optogenetics, photoproximity labeling, protein-protein interactions",
author = "Nir Hananya and Xuanjia Ye and Shany Koren and Muir, {Tom W.}",
note = "Funding Information: ACKNOWLEDGMENTS. We thank members of the Muir laboratory for valuable discussions and comments. We thank Saw Kyin and Henry H. Shwe from the Proteomics and Mass Spectrometry Core Facility, Gary Laevsky and Sha Wang from the Confocal Imaging Facility, and Christina J. DeCoste and Katherine Rittenbach from the Flow Cytometry Resource Facility at Princeton University. We thank Dr. Steve Roffler for his helpful advice regarding the expression of LOV* on the cell surface. This work was supported by grants from the NIH [(R37-GM0968`68 and P01-CA196539 to T.W.M.]. N.H. is a Robert Black Fellow of the Damon Runyon Cancer Research Foundation, DRG-2425-21. X.Y. is supported by a graduate fellowship from the China Scholarship Council. S.K. is supported by Human Frontier Science Program fellowship, LT000595/2020 and was supported by a Swiss National Science Foundation Postdoc.Mobility Fellowship (P400PB_191056). Funding Information: We thank members of the Muir laboratory for valuable discussions and comments. We thank Saw Kyin and Henry H. Shwe from the Proteomics and Mass Spectrometry Core Facility, Gary Laevsky and Sha Wang from the Confocal Imaging Facility, and Christina J. DeCoste and Katherine Rittenbach from the Flow Cytometry Resource Facility at Princeton University. We thank Dr. Steve Roffler for his helpful advice regarding the expression of LOV* on the cell surface. This work was supported by grants from the NIH [(R37-GM096868 and P01-CA196539 to T.W.M.]. N.H. is a Robert Black Fellow of the Damon Runyon Cancer Research Foundation, DRG-2425-21. X.Y. is supported by a graduate fellowship from the China Scholarship Council. S.K. is supported by Human Frontier Science Program fellowship, LT000595/2020 and was supported by a Swiss National Science Foundation Postdoc.Mobility Fellowship (P400PB_191056). Publisher Copyright: Copyright {\textcopyright} 2023 the Author(s). Published by PNAS. This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).",
year = "2023",
month = apr,
day = "18",
doi = "10.1073/pnas.2219339120",
language = "English (US)",
volume = "120",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "16",
}