title: Use of the arginine-specific butanedione/phenylboronic acid tag for analysis of peptides and protein digests using matrix-assisted laser desorption/ionization mass spectrometry.
abstract: 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 alpha-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.
authors.affiliation: Department of Analytical Chemistry and Food Chemistry, University of Vienna, Waehringer Strasse 38, 1090 Vienna, Austria. alexander.leitner@univie.ac.at
authors.forename: Alexander
authors.initials: A
authors.lastname: Leitner
authors.forename: Sabine
authors.initials: S
authors.lastname: Amon
authors.forename: Andreas
authors.initials: A
authors.lastname: Rizzi
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lindner
mesh_terms: D001120:Arginine / Q000737:chemistry*; D001897:Boronic Acids / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D001897:Boronic Acids; D004852:Epoxy Compounds; D010455:Peptides; C007366:diepoxybutane; D001120:Arginine; D010447:Peptide Hydrolases; C010686:benzeneboronic acid