title: C-terminal sequence determination of peptides degraded with carboxypeptidases of different specificities and analyzed by 252-Cf plasma desorption mass spectrometry.
abstract: Identification of the truncated peptides by plasma desorption mass spectrometry in C-terminal sequence determination with carboxypeptidases offers several advantages over analysis of the liberated amino acids. It is possible to perform in situ digestion of a nitrocellulose-bound sample already used for molecular weight determination and thus obtain sequence information without further sample consumption. In time-course analysis the analytical information, although not obtained in real time, is sufficiently rapid to adjust the digestion conditions. There is no need for quantitation because the identification is based on molecular weight differences. Sensitivity in the low picomole range is obtainable. The digestion of a number of peptides (900-3500 Da) with carboxypeptidase Y and MII has been monitored. It was found that successive use of the enzymes or use of a mixture of the enzymes was often advantageous. The sequence of up to 10 residues from the C-terminus has been determined for the peptides studied.
authors.affiliation: Department of Molecular Biology, Odense University, Denmark.
authors.forename: I
authors.initials: I
authors.lastname: Klarskov
authors.forename: K
authors.initials: K
authors.lastname: Breddam
authors.forename: P
authors.initials: P
authors.lastname: Roepstorff
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002268:Carboxypeptidases; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D007328:Insulin / Q000031:analogs & derivatives; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis*; D010940:Plant Proteins; D012259:Ribonuclease, Pancreatic; D012996:Solutions; D014357:Trypsin
chemical_list: D007328:Insulin; D010446:Peptide Fragments; D010940:Plant Proteins; D012996:Solutions; D012259:Ribonuclease, Pancreatic; D002268:Carboxypeptidases; C022633:serine carboxypeptidase; C061521:CP-MII protein, Hordeum vulgare; D002918:Chymotrypsin; D014357:Trypsin