Isolation and identification of C-terminal peptides of proteins.
A method for the isolation and identification of C-terminal peptides of proteins has been developed. The procedure entails the racemization of the C-terminal amino acid by reaction of the N-trifluoroacetylated protein with acetic anhydride and pyridine. After deprotection, the protein is fragmented to yield a mixture of peptides, which in turn are digested with carboxypeptidases. All peptides are hydrolyzed to L-amino acids except the C-terminal peptide with the terminal D-amino-acid residue. It is resistant to the action of carboxypeptidases and is readily identified by peptide mapping.
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School of Pharmacy, University of Wisconsin, Madison, USA.
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D000089:Acetic Anhydrides / Q000737:chemistry; D000595:Amino Acid Sequence; D001920:Bradykinin / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D002268:Carboxypeptidases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009496:Neurotensin / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry*; D011725:Pyridines / Q000737:chemistry; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014867:Water / Q000737:chemistry
D000089:Acetic Anhydrides; D010446:Peptide Fragments; D011506:Proteins; D011725:Pyridines; D014867:Water; C031800:acetic anhydride; D009496:Neurotensin; D014269:Trifluoroacetic Acid; D002268:Carboxypeptidases; D002256:Carbonic Anhydrases; C023666:pyridine; D001920:Bradykinin