Identification of cysteine-containing peptides in protein digests by high-performance liquid chromatography.
A method is described for the facile identification of half-cystine residues in peptides from protein digests. Combined use of S-beta-(4-pyridylethylation) of cysteine residues with 4-vinyl pyridine and peptide mapping by reversed-phase high-performance liquid chromatography provides the basis for this procedure. Essentially simultaneous, in-line effluent monitoring at 220, 280, and 254 nm provides highly sensitive, specific, and instantaneous identification of all peptide peaks, as well as those containing tryptophan and half-cystine residues, respectively. The resultant cysteine-derivatized peptides are well suited to amino acid compositional and protein sequence analysis, and are protected from loss during enzymatic digestion and/or peptide mapping.
D000818:Animals; D002135:Calcium-Binding Proteins / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000031:analogs & derivatives / Q000032:analysis*; D006868:Hydrolysis; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*
D002135:Calcium-Binding Proteins; D010455:Peptides; D011506:Proteins; C009127:S-beta-(4-pyridylethyl)-L-cysteine; D003545:Cysteine