title: Sequencing of peptides and proteins from the carboxy terminus.
abstract: A new chemical method for carboxy-terminal (C-terminal) protein sequencing has been developed. This approach has been successfully used to sequence 5 residues of standard proteins and 5 to 10 residues of synthetic peptides at low nanomole levels. The sequencing procedure consists of converting the C-terminal amino acid into a thiohydantoin (TH) derivative, followed by transformation of the TH into a good leaving group by alkylation. Next, the alkylated TH is cleaved mildly and efficiently with (N = C V S)- anion, which simultaneously forms a TH on the newly truncated protein or peptide. Thus, after the initial TH derivatization, there is no return to a free carboxyl group at the C-terminus. An additional benefit of this method is that the alkylating moiety can be chosen with a variety of properties allowing for variation in the detection method. This chemistry has been adapted to automated protein sequencers with a cycle time of about 1 h.
authors.affiliation: Applied Biosystems, Inc., Foster City, California 94404.
authors.forename: V L
authors.initials: VL
authors.lastname: Boyd
authors.forename: M
authors.initials: M
authors.lastname: Bozzini
authors.forename: G
authors.initials: G
authors.lastname: Zon
authors.forename: R L
authors.initials: RL
authors.lastname: Noble
authors.forename: R J
authors.initials: RJ
authors.lastname: Mattaliano
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
chemical_list: D007202:Indicators and Reagents; D010455:Peptides; D011506:Proteins