Huang I Y, Welch C D, Yoshida A
J Biol Chem. 1980 Jul 10;255(13):6412-20.
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.
用溴化氰处理还原的S - 羧甲基化磷酸甘油酸激酶产生了14个主要肽段,即CNBr - 1(20个残基)、CNBr - 2(8个残基)、CNBr - 3(33个残基)、CNBr - 4(11个残基)、CNBr - 5(104个残基)、CNBr - 6(14个残基)、CNBr - 7(37个残基)、CNBr - 8(7个残基)、CNBr - 9(6个残基)、CNBr - 10(11个残基)、CNBr - 11(19个残基)、CNBr - 12(42个残基)、CNBr - 13(44个残基)和CNBr - 14(61个残基)。通过自动和手动序列分析相结合,以及对胰蛋白酶肽段、糜蛋白酶肽段和经葡萄球菌蛋白酶消化获得的肽段进行表征,确定了所有溴化氰肽段的氨基酸序列。从溴化氰肽段CNBr - 13和CNBr - 14中回收了两个通过直接消化完整磷酸甘油酸激酶未获得的胰蛋白酶肽段,并对这些肽段进行了纯化和测序。基于从该酶衍生的所有胰蛋白酶和溴化氰肽段的信息,对这些肽段进行了正确的比对。因此,确定了由417个氨基酸残基组成的人磷酸甘油酸激酶的完整氨基酸序列。