Improved collision-induced dissociation analysis of peptides by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry through 3-sulfobenzoic acid succinimidyl ester labeling.
The sulfonation reagent, a succinimidyl ester of 3-sulfobenzoic acid, has been synthesized for effective peptide sequencing. It is capable of incorporating an additional mobile proton into the peptide backbone, thus, facilitating efficient collision-induced dissociation. This reagent is easily and inexpensively prepared in short time. Tandem mass spectra of the guanidinated and reagent-sulfonated peptides consist mainly of the y-ion series with higher intensities than those observed for solely guanidinated peptides. These enhanced tandem MS attributes significantly improved MASCOT total-ion scores, thus, allowing more confident peptide sequencing. This derivatization was also very effective for the analysis of tryptic digest of human blood serum proteins separated by two-dimensional gel electrophoresis. When used in LC-MALDI/MS/MS format, this type of derivatization does not adversely affect chromatographic efficiencies.
Alley
William R
WR
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Mechref
Yehia
Y
Klouckova
Iveta
I
Novotny
Milos V
MV
D000595:Amino Acid Sequence; D000818:Animals; D016632:Apolipoprotein A-I / Q000737:chemistry; D001557:Benzenesulfonates / Q000737:chemistry / Q000494:pharmacology*; D001798:Blood Proteins / Q000737:chemistry*; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004952:Esters / Q000737:chemistry*; D006736:Horses / Q000378:metabolism; D006801:Humans; D007834:Lasers; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
C521207:3-sulfobenzoic acid succinimidyl ester; D016632:Apolipoprotein A-I; D001557:Benzenesulfonates; D001798:Blood Proteins; D004952:Esters; D009211:Myoglobin; D010455:Peptides