title: Analyses of protein-protein interactions by in vivo photocrosslinking in budding yeast.
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.
authors.affiliation: Department of Chemistry, Nagoya University, Nagoya, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Shiota
authors.forename: Shuh-ichi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Toshiya
authors.initials: T
authors.lastname: Endo
mesh_terms: D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D033681:Mitochondrial Membrane Transport Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D055668:Photochemical Processes; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: D005656:Fungal Proteins; D033681:Mitochondrial Membrane Transport Proteins