title: Identification of proteins based on MS/MS spectra and location of posttranslational modifications.
abstract: Comparative protein profiling is a key approach to understanding the human and other proteomes. Systems-level profiling technologies, such as differential fluorescence two-dimensional gel electrophoresis (DIGE), often require the identification of the proteins that are contained within 50 or more spots per gel. A major focus of this chapter therefore is devoted to a general approach for high throughput protein identification that is based on liquid chromatography (LC)/tandem mass spectrometry (MS/MS) analysis of tryptic digests of individual proteins or mixtures of only a few proteins (i.e., as are usually obtained from individual DIGE spots), and that is also applicable to the analysis of complex protein extracts. Additionally, multiple techniques will be described for identifying sites of protein posttranslational modification, with emphasis on phosphorylation and Arg methylation.
authors.affiliation: WM Keck Foundation Biotechnology Resource Laboratory, Mass Spectrometry and Protomic Resources, Yale University, New Haven, CT, USA.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Stone
authors.forename: Myron
authors.initials: M
authors.lastname: Crawford
authors.forename: Walter
authors.initials: W
authors.lastname: McMurray
authors.forename: Nancy
authors.initials: N
authors.lastname: Williams
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Williams
mesh_terms: D000469:Alkaline Phosphatase; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008967:Molecular Biology / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping / Q000379:methods; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin
chemical_list: D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D011506:Proteins; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D000469:Alkaline Phosphatase; D014357:Trypsin