Complete amino acid sequence of human phosphoglycerate kinase. Cyanogen bromide peptides and complete amino acid sequence

Abstract

Cyanogen bromide treatment of reduced, S-carboxymethylated phosphoglycerate kinase yielded 14 major peptides, CNBr-1 (20 residues), CNBr-2 (8 residues), CNBr-3 (33 residues), CNBr-4 (11 residues), CNBr-5 (104 residues), CNBr-6 (14 residues), CNBr-7 (37 residues), CNBr-8 (7 residues), CNBr-9 (6 residues), CNBr-10 (11 residues), CNBr-11 (19 residues), CNBr-12 (42 residues), CNBr-13 (44 residues), and CNBr-14 (61 residues). The amino acid sequences of all the cyanogen bromide peptides were determined by a combination of automated and manual sequence analysis, and the characterization of tryptic and chymotryptic peptides and peptides obtained by digestion with staphylococcal protease. Two tryptic peptides which were not obtained by direct digestion of whole phosphoglycerate kinase were recovered from cyanogen bromide Peptides CNBr-13 and CNBr-14 and these peptides were purified and sequenced. Based on the information from all the tryptic and cyanogen bromide peptides derived from the enzyme, the proper alignment of these peptides was made. Thus, complete amino acid sequence of human phosphoglycerate kinase consisting of 417 amino acid residues was determined.