Protein sequencing by tandem mass spectrometry.

Abstract

Methodology for determining amino acid sequences of proteins by tandem mass spectrometry is described. The approach involves enzymatic and/or chemical degradation of the protein to a collection of peptides which are then fractionated by high-performance liquid chromatography. Each fraction, containing as many as 10-15 peptides, is then analyzed directly, without further purification, by a combination of liquid secondary-ion/collision-activated dissociation mass spectrometry on a multianalyzer instrument. Interpretation of collision-activated dissociation mass spectra is described, and results are presented from a study of soluble peptides produced by treatment of apolipoprotein B with cyanogen bromide and trypsin.

Keywords

ChemistryCyanogen bromideMass spectrometryTandem mass spectrometryChromatographyTandem mass tagProtein mass spectrometryCollision-induced dissociationBottom-up proteomicsTop-down proteomicsTrypsinSample preparation in mass spectrometryIsobaric labelingPeptideCyanogenDissociation (chemistry)Peptide sequenceProteomicsBiochemistryEnzymeOrganic chemistryQuantitative proteomicsElectrospray ionization

Affiliated Institutions

University of Virginia US

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Publication Info

Year: 1986
Type: article
Volume: 83
Issue: 17
Pages: 6233-6237
Citations: 1172
Access: Closed

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APA Style

                            
                                
                                    Donald F. Hunt, 
                                
                                    John R. Yates, 
                                
                                    Jeffrey Shabanowitz
                                
                                et al.
                            
                            (1986). 
                            Protein sequencing by tandem mass spectrometry.. 
                            Proceedings of the National Academy of Sciences
                            , 83
                            (17)
                            , 6233-6237.
                            https://doi.org/10.1073/pnas.83.17.6233
                        

Identifiers

DOI: 10.1073/pnas.83.17.6233