Use of immobilized exopeptidases and volatile buffers for analysis of peptides by fast atom bombardment mass spectrometry.
beta-Lipotrophin (62-77) or Ac-gastrin releasing peptide was incubated with immobilized carboxypeptidase Y or aminopeptidase M. Subsequent aliquots of each incubation mixture were analysed by fast atom bombardment mass spectrometry using a dithiothreitol/dithioerythritol liquid matrix. The use of immobilized enzymes and volatile buffers for exopeptidase digestions enabled rapid and facile separation of enzyme from digestion products. This approach to mass spectral peptide analysis reduced spectral background arising from a glycerol matrix, buffer salts, or enzyme proteins and contaminants, enabling analysis of as little as 200 picomoles of a suitable peptide.
D000595:Amino Acid Sequence; D000626:Aminopeptidases; D002021:Buffers; D018826:CD13 Antigens; D002268:Carboxypeptidases; D004800:Enzymes, Immobilized; D013058:Mass Spectrometry / Q000379:methods; D010446:Peptide Fragments / Q000032:analysis*; D008083:beta-Lipotropin / Q000032:analysis*
D002021:Buffers; D004800:Enzymes, Immobilized; D010446:Peptide Fragments; D008083:beta-Lipotropin; D002268:Carboxypeptidases; D000626:Aminopeptidases; D018826:CD13 Antigens; C022633:serine carboxypeptidase