title: Identification of nearest-neighbor peptides in protease digests by mass spectrometry for construction of sequence-ordered tryptic maps.
abstract: Continuous-flow fast atom bombardment mass spectrometry was used for the identification of the intermediates and end products of the tryptic digest of polypeptides throughout the time-course of the reactions. Precursor/product relationships for these peptides were determined with the aid of a simple personal computer program. The C-terminal tryptic peptides were identified by performing a tryptic digest in 50% 18O-enriched buffer which resulted in labeling of all non-C-terminal peptides with 18O. This information, along with the precursor/product correlations, was used to create a sequence-ordered tryptic map of the original polypeptide. Ion intensities of intermediate hydrolysis products are compared for enzyme to substrate ratios of 1:100 and 1:1000 (w/w) over the course of the reaction. Intermediates were found to have significantly longer lifetimes when lower levels of trypsin were present.
authors.affiliation: Analytical Chemistry Center, University of Texas Medical School, Houston 77225.
authors.forename: B
authors.initials: B
authors.lastname: Whaley
authors.forename: R M
authors.initials: RM
authors.lastname: Caprioli
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002116:Calcitonin; D004524:Eels; D006801:Humans; D008969:Molecular Sequence Data; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D010449:Peptide Mapping / Q000379:methods*; D051381:Rats; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods; D019379:Teriparatide; D014357:Trypsin
chemical_list: D010281:Parathyroid Hormone; D010446:Peptide Fragments; D019379:Teriparatide; D002116:Calcitonin; D014357:Trypsin