Shoji S, Ericsson L H, Walsh K A, Fischer E H, Titani K
Biochemistry. 1983 Jul 19;22(15):3702-9. doi: 10.1021/bi00284a025.
The 350-residue amino acid sequence of the catalytic subunit of bovine cardiac muscle adenosine cyclic 3',5'-phosphate dependent protein kinase is described. The protein has a molecular weight of 40 862, which includes an N-tetradecanoyl (myristyl) group blocking the NH2 terminus and phosphate groups at threonine-197 and serine-338. Seven methionyl bonds in the S-carboxymethylated protein were cleaved with cyanogen bromide to yield eight primary peptides. These fragments, and subpeptides generated by cleavage with trypsin, pepsin, chymotrypsin, thermolysin, and Myxobacter AL-1 protease II, were purified and analyzed to yield the majority of the sequence. The primary peptides were aligned by analyses of overlapping peptides, particularly of methione-containing tryptic peptides generated after in vitro [14C]methyl exchange labeling of methionyl residues in the intact protein.
本文描述了牛心肌腺苷环化3',5'-磷酸依赖性蛋白激酶催化亚基的350个氨基酸残基序列。该蛋白分子量为40862，其氨基端被N-十四烷酰基（肉豆蔻酰基）封闭，在苏氨酸-197和丝氨酸-338处有磷酸基团。用溴化氰裂解S-羧甲基化蛋白中的七个甲硫氨酸键，产生八个一级肽段。这些片段以及用胰蛋白酶、胃蛋白酶、胰凝乳蛋白酶、嗜热菌蛋白酶和粘细菌AL-1蛋白酶II裂解产生的亚肽段经过纯化和分析，得出了大部分序列。通过分析重叠肽段，特别是完整蛋白中甲硫氨酸残基经体外[14C]甲基交换标记后产生的含甲硫氨酸的胰蛋白酶肽段，对一级肽段进行了比对。
