Premium
Peptide fingerprints after partial acid hydrolysis: Analysis by matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Knierman Michael D.,
Coligan John E.,
Parker Kenneth C.,
Chait B. T.
Publication year - 1994
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/rcm.1290081220
Subject(s) - chemistry , peptide , mass spectrometry , hydrolysate , hydrolysis , chromatography , peptide sequence , matrix assisted laser desorption/ionization , protein mass spectrometry , amino acid , desorption , acid hydrolysis , hydrochloric acid , peptide mass fingerprinting , matrix (chemical analysis) , tandem mass spectrometry , biochemistry , organic chemistry , proteomics , adsorption , gene
Abstract A method is described by which a sequence‐dependent peptide fingerprint can be rapidly obtained upon partial hydrolysis of peptides with hydrochloric acid and subsequent analysis by matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS). When synthetic peptides are treated with 3M HCI for 5 min at 110°C, amino acids are released in turn from the C‐terminus or, depending on the peptide, from the N‐terminus. Sequence information can be deduced by identifying the amino acid whose mass corresponds to the difference in MW between the major hydrolysis products, beginning from the MW of the starting peptide. A similar pattern exclusively from the C‐terminus has been obtained using pentafluoropropionic acid as a hydrolyzing agent (Tsugita et al. Eur. J. Biochem. 206, 691 (1992)), but required longer hydrolysis time and more handling prior to analysis. The technique we have developed could be used to obtain a sequence‐dependent ‘fingerprint’ for a peptide cheaply and rapidly, starting with picomole amounts of peptide, because the hydrolysate can be directly analyzed by MALDI. This methodology might be especially useful for confirming the identity of peptides during peptide mapping.