Identification of proteins based on MS/MS spectra and location of posttranslational modifications.
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.
Stone
Kathryn L
KL
WM Keck Foundation Biotechnology Resource Laboratory, Mass Spectrometry and Protomic Resources, Yale University, New Haven, CT, USA.
Crawford
Myron
M
McMurray
Walter
W
Williams
Nancy
N
Williams
Kenneth R
KR
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
D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D011506:Proteins; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D000469:Alkaline Phosphatase; D014357:Trypsin