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In Vivo Photocrosslinking with Unnatural Amino Acid Mutagenesis
Author(s) -
Chin Jason W.,
Schultz Peter G.
Publication year - 2002
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20021104)3:11<1135::aid-cbic1135>3.0.co;2-m
Subject(s) - mutagenesis , amino acid , covalent bond , in vivo , chemistry , monomer , biochemistry , intracellular , biophysics , biology , mutation , genetics , gene , organic chemistry , polymer
Shining a light on cells allows an intracellular protein to be covalently crosslinked to its in vivo binding partner. Unnatural amino acid mutagenesis is used to introduce a photocrosslinking amino acid into selected sites in monomers of a dimeric complex. In vivo surface‐specific photocrosslinking of these monomers (see figure) is demonstrated by purification of the complexes formed and their identification by Western blot. This methodology may be used to elucidate the many varied interactions of proteins within the cell.