Analyses of protein-protein interactions by in vivo photocrosslinking in budding yeast

Takuya Shiota, Shuh Ichi Nishikawa, Toshiya Endo

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

10 Citations (Scopus)

Abstract

Recent development of methods for genetic incorporation of unnatural amino acids into proteins in live cells enables us to analyze protein interactions by site-specific photocrosslinking. Here we describe a method to incorporate p-benzoyl-l-phenylalanine (pBpa), a photoreactive unnatural amino acid, into defined positions of a target protein in living yeast cells. Photocrosslinking using the pBpa-incorporated proteins has been proven to be a powerful method for analyzing protein-protein interactions at the spatial resolution of amino-acid residues. Since photocrosslinking can be performed for pBpa-incorporated proteins that are properly assembled into a protein complex in living cells, this method will allow us to reveal protein-protein interactions of the target proteins at work.

Original languageEnglish
Title of host publicationMembrane Biogenesis: Methods and Protocols
EditorsDoron Rapaport, Johannes M. Herrmann
Place of PublicationNew York NY USA
PublisherSpringer Science + Business Media
Chapter14
Pages207-217
Number of pages11
Volume1033
ISBN (Print)9781627034869
DOIs
Publication statusPublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1033
ISSN (Print)10643745

Keywords

  • Photocrosslink
  • Protein interactions
  • Suppressor tRNA
  • Unnatural amino acids
  • Yeast

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