Peptide sequence analysis using exopeptidases with molecular analysis of the truncated polypeptides by mass spectrometry.

Fast atom bombardment mass spectrometry is used for the analysis of the series of molecular products formed by the cleavage of polypeptide substrates with the exopeptidases carboxypeptidase Y and leucine aminopeptidase. By following the polypeptide molecular species rather than the released residues, sequence information is obtained regardless of the relative rates of cleavage of peptide bonds. In addition, unambiguous assignments of sequence can be made in the presence of multiple identical residues. The lower level of sensitivity for the analysis is in the picomole range. When carboxypeptidase Y is used, the method provides a specific and sensitive method for the sequencing of polypeptides from the C-terminus.