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Asparaginyl endopeptidase mapping of proteins with subsequent matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Kanda Fujihiro,
Yoshida Seiichi,
Okumura Tatsuya,
Takamatsu Tasuku
Publication year - 1995
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.1290091203
Subject(s) - chemistry , mass spectrometry , chromatography , matrix assisted laser desorption/ionization , endopeptidase , protein mass spectrometry , sample preparation in mass spectrometry , bottom up proteomics , matrix (chemical analysis) , sample preparation , proteolysis , desorption , electrospray ionization , enzyme , biochemistry , organic chemistry , adsorption
A matrix‐assisted laser desorption/ionization (MALDI) mass spectrometric mapping strategy for the identification and characterization of isolated and purified proteins is described. The method, which employs the combined usage of a new site‐specific enzyme Asparaginyl endopeptidase (Asn‐EP) for proteolysis, and MALDI for subsequent mass analysis, is capable of rapidly and sensitively examining the components of complex mixtures without any chromatographic or electrophoretic separation steps. Subpicomole sample quantities typically suffice to permit the confirmation of deduced primary structures and/or the identification of possible post‐translational modifications. The data obtained should also prove useful for mass matching and sequence homology searching of computerised protein sequence data bases of known proteins.