title: Mass spectrometric determination of protein ubiquitination.
abstract: Mass spectrometric methods of determining protein ubiquitination are described. Characteristic mass shifts and fragment ions indicating ubiquitinated lysine residues in tryptic and gluC digests are discussed. When a ubiquitinated protein is enzymatically digested, a portion of the ubiquitin side chain remains attached to the modified lysine. The ubiquitinated peptide thus has two N-termini- one from the original peptide and one from the ubiquitin side chain. Thus, it is possible to have two series of b ions and y ions, the additional series is the one that includes fragments containing portions of the ubiquitin side chain. Any diagnostic ions for the modification must include portions of this side chain. Fragment ions involving any part of the "normal" peptide will vary in mass according to the peptide being modified and will therefore not be of general diagnostic use. These diagnostic ions, found through examination of the MS/MS spectra of model ubiquitinated tryptic and gluC peptides, can be used to trigger precursor ion scanning in automated MS/MS data acquisition scanning modes.
authors.affiliation: Department of Biochemistry and Biophysics and the Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, USA.
authors.forename: Carol E
authors.initials: CE
authors.lastname: Parker
authors.forename: Viorel
authors.initials: V
authors.lastname: Mocanu
authors.forename: Maria R
authors.initials: MR
authors.lastname: Warren
authors.forename: Susanna F
authors.initials: SF
authors.lastname: Greer
authors.forename: Christoph H
authors.initials: CH
authors.lastname: Borchers
mesh_terms: D000818:Animals; D006801:Humans; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D025801:Ubiquitin / Q000737:chemistry*
chemical_list: D025801:Ubiquitin