A genetically encoded photoproximity labeling approach for mapping protein territories.

Autor:	Hananya N; Department of Chemistry, Princeton University, Princeton, NJ 08544., Ye X; Department of Chemistry, Princeton University, Princeton, NJ 08544., Koren S; Department of Chemistry, Princeton University, Princeton, NJ 08544., Muir TW; Department of Chemistry, Princeton University, Princeton, NJ 08544.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2023 Apr 18; Vol. 120 (16), pp. e2219339120. Date of Electronic Publication: 2023 Apr 10.
DOI:	10.1073/pnas.2219339120
Abstrakt:	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.
Databáze:	MEDLINE
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