Selective identification and quantitative analysis of methionine containing peptides by charge derivatization and tandem mass spectrometry
Details
Publication Year 2005-07, Volume 16, Issue #7, Page 1131-1150
Journal Title
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Publication Type
Journal Article
Abstract
To enable the development of a tandem mass spectrometry (MS/MS) based methodology for selective protein identification and differential quantitative analysis, a novel derivatization strategy is proposed, based on the formation of a "fixed-charge" sulfonium ion on the side-chain of a methionine amino acid residue contained within a protein or peptide of interest. The gas-phase fragmentation behavior of these side chain fixed charge sulfonium ion containing peptides is observed to result in exclusive loss of the derivatized side chain and the formation of a single characteristic product ion, independently of charge state or amino acid composition. Thus, fixed charge containing peptide ions may be selectively identified from complex mixtures, for example, by selective neutral loss scan mode MS/MS methods. Further structural interrogation of identified peptide ions may be achieved by subjecting the characteristic MS/MS product ion to multistage MS/MS (MS3) in a quadrupole ion trap mass spectrometer, or by energy resolved "pseudo" MS3 in a triple quadrupole mass spectrometer. The general principles underlying this fixed charge derivatization approach are demonstrated here by MS/MS, MS3 and "pseudo" MS3 analysis of side chain fixed-charge sulfonium. ion derivatives of peptides containing methionine formed by reaction with phenacylbromide. Incorporation of "light" and "heavy" isotopically encoded labels into the fixed-charge derivatives facilitates the application of this method to the quantitative analysis of differential protein expression, via measurement of the relative abundances of the neutral loss product ions generated by dissociation of the light and heavy labeled peptide ions. This approach, termed "selective extraction of labeled entities by charge derivatization and tandem mass spectrometry" (SELECT), thereby offers the potential for significantly improved sensitivity and selectivity for the identification and quantitative analysis of peptides or proteins containing selected structural features, without requirement for extensive fractionation or otherwise enrichment from a complex mixture prior to analysis. (c) 2005 American Society for Mass Spectrometry
Publisher
ELSEVIER SCIENCE INC
Keywords
COMPLEX PROTEIN MIXTURES; POSITIVE-ION MODE; COMPARATIVE PROTEOMICS; AFFINITY TAG; PHOSPHORYLATED PEPTIDES; GEL-ELECTROPHORESIS; NEIGHBORING GROUP; SULFONIUM SALTS; ACTIVE-SITE; AMINO-ACIDS
Rights Notice
Refer to copyright notice on published article.


Creation Date: 2005-07-01 12:00:00
Last Modified: 0001-01-01 12:00:00
An error has occurred. This application may no longer respond until reloaded. Reload 🗙