Monitoring of Carboxypeptidase Digestion by Matrix-Assisted Laser Desorption and Ionization Mass Spectrometry

Abstract

The potential of matrix-assisted laser desorption and ionization mass spectrometry (LDI-MS) is demonstrated by monitoring and analyzing the digestion of (human) pTH (1–34), a synthetic peptide with carboxypeptidases Y and B. All occurring ion signals in the mass spectra could be identified as degraded peptides. By calculating the mass differences between successive degraded peptides, it was possible to identify the released amino acids and to determine 8 amino acids of the C-terminus of the original peptide. For a single MS measurement, only 2 pmol of substrate was needed. Time-course analysis of the cleavage of the first amino acid residue gave insight into the kinetics involved. These measurements strongly support the hope that quantitative information about concentrations can be extracted from LDI-MS.