An isotope coding strategy for proteomics involving both amine and carboxyl group labeling.
A stable isotope coding strategy is described for the analysis of all types of tryptic peptides, including those that are N-terminally blocked and from the C-terminus of proteins. The method exploits differential derivatization of amine and carboxyl groups generated during proteolysis as a means of coding. Carboxyl groups produced during proteolysis incorporate 18O from H2(18)O. Peptides from the C-terminus of proteins were not labeled with 18O unless they contained a basic C-terminal amino acid. Primary amines form controls, and experimental samples were differentially acylated after proteolysis with either 1H3- or 2H3-N-acetoxysuccinamide. When these two types of labeling were combined, unique coding patterns were achieved for peptides arising from the C-termini and blocked N-termini of proteins.
Regnier
Fred E
FE
Department of Chemistry and Cancer Center, Purdue University, West Lafayette, IN, USA.
D000588:Amines / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods*; D015394:Molecular Structure; D010103:Oxygen Isotopes / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D000588:Amines; D010103:Oxygen Isotopes; D010455:Peptides