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A strategy for rapid and efficient sequencing of Lys‐C peptides by matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry post‐source decay
Author(s) -
Pfeifer Thomas,
Rücknagel Peter,
Kuellertz Gerhard,
Schierhorn Angelika
Publication year - 1999
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19990315)13:5<362::aid-rcm492>3.0.co;2-i
Subject(s) - chemistry , mass spectrometry , matrix assisted laser desorption/ionization , peptide , amino acid , acetylation , peptide sequence , time of flight mass spectrometry , matrix (chemical analysis) , lysine , chromatography , ion , desorption , biochemistry , ionization , organic chemistry , adsorption , gene
A modification procedure for Lys‐C peptides is described which simplifies the correct assignment of the amino acid sequence. Release of the C‐terminal lysine from Lys‐C peptides by carboxypeptidase B and subsequent N‐terminal acetylation of the resulting peptides leads to predictable shifts of the C‐ and N‐terminal fragment ions in Matrix‐assisted laser desorption/ionisation time‐of‐flight post‐source decay mass spectra and facilitates the correct assignment of mostly complete amino acid sequences for oligopeptides. The derived sequences of peptides from unknown proteins were used to search in databases for homologous protein sequences. Our method was applied to an unknown protein isolated from eggs of Drosophila melanogaster , resulting in the identification of a peptidyl prolyl cis‐trans ‐isomerase. Copyright © 1999 John Wiley & Sons, Ltd.