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Microwave‐enhanced enzyme reaction for protein mapping by mass spectrometry: A new approach to protein digestion in minutes
Author(s) -
Pramanik Birendra N.,
Mirza Urooj A.,
Ing Yao Hain,
Liu YanHui,
Bartner Peter L.,
Weber Patricia C.,
Bose Ajay K.
Publication year - 2002
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.0213702
Subject(s) - chemistry , trypsin , mass spectrometry , peptide , lysozyme , myoglobin , chromatography , fragmentation (computing) , matrix assisted laser desorption/ionization , proteolysis , electrospray ionization , peptide mapping , sample preparation in mass spectrometry , lysine , biochemistry , enzyme , peptide sequence , desorption , biology , amino acid , ecology , organic chemistry , adsorption , gene
Accelerated proteolytic cleavage of proteins under controlled microwave irradiation has been achieved. Selective peptide fragmentation by endoproteases trypsin or lysine C led to smaller peptides that were analyzed by matrix‐assisted laser desorption ionization (MALDI) or liquid chromatography‐electrospray ionization (LC‐ESI) techniques. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analyses of the peptide fragmentation of several biologically active proteins, including cytochrome c, ubiquitin, lysozyme, myoglobin, and interferon α‐2b. Most important, using this novel approach digestion of proteins occurs in minutes, in contrast to the hours required by conventional methods.