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Updated: May 8, 2026

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos
Published on: April 23, 2014
Analyses of protein-protein interactions by in vivo photocrosslinking in budding yeast
Takuya Shiota1, Shuh-ichi Nishikawa, Toshiya Endo
1Department of Chemistry, Nagoya University, Nagoya, Japan.
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.
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