A genetically encoded photoproximity labeling approach for mapping protein territories

Nir Hananya, Xuanjia Ye, Shany Koren, Tom W. Muir

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

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.

Original languageEnglish (US)
Article numbere2219339120
JournalProceedings of the National Academy of Sciences of the United States of America
Volume120
Issue number16
DOIs
StatePublished - Apr 18 2023

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • LOV domain
  • optogenetics
  • photoproximity labeling
  • protein-protein interactions

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