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Use of the arginine‐specific butanedione/phenylboronic acid tag for analysis of peptides and protein digests using matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Leitner Alexander,
Amon Sabine,
Rizzi Andreas,
Lindner Wolfgang
Publication year - 2007
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.2967
Subject(s) - chemistry , chromatography , mass spectrometry , protein mass spectrometry , matrix assisted laser desorption/ionization , sample preparation in mass spectrometry , peptide , arginine , desorption , amino acid , tandem mass spectrometry , electrospray ionization , biochemistry , organic chemistry , adsorption
We have applied an arginine‐specific labeling technique to the study of peptides by matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS). The reaction converts the guanidine group of the arginine side chain by reacting it with 2,3‐butanedione and an arylboronic acid. Despite the general chemical lability of the tag under acidic conditions, it was possible to employ acidic matrices like α ‐cyano‐4‐hydroxycinnamic acid without adverse effects, using the thin‐layer technique for preparation. After optimizing the method using arginine‐containing model peptides – for which sensitivity down to the low fmol range was demonstrated – the procedure was applied to enzymatic digests of several model proteins in solution and to protein spots in gels obtained by two‐dimensional electrophoretic separation of cell lysate samples. Information on the presence of arginine in peptides can be easily obtained from the mass spectra by the characteristic mass shift and the isotope pattern resulting from the incorporation of boron. This information might serve as a valuable additional search constraint for achieving a higher degree of confidence for protein identification by peptide mass fingerprinting. Copyright © 2007 John Wiley & Sons, Ltd.