Identification of components of protein complexes.
Protocols are given for a variety of techniques used in protein identification of complexes, including identification of in-gel separated proteins and LC-MS/MS. Gels, staining procedures, and peptide extraction protocols that are compatible with mass spectrometry are described. The detection limits of the various staining procedures and their compatibility with mass spectrometry are discussed. The various mass spectrometric techniques used (MALDI-MS, MALDI-MS/MS, nanospray, and ESI/LC-MS/MS) are also described, along with an indication of the advantages and disadvantages of each, and when they would most appropriately be used. Common pitfalls associated with database searching are also discussed.
Parker
Carol E
CE
Department of Biochemistry and Biophysics and the Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, USA.
Warren
Maria R
MR
Loiselle
David R
DR
Dicheva
Nedyalka N
NN
Scarlett
Cameron O
CO
Borchers
Christoph H
CH
D000818:Animals; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D010449:Peptide Mapping / Q000379:methods; D040901:Proteomics / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013194:Staining and Labeling