Complete amino acid sequence of pig kidney fructose-1,6-bisphosphatase.
Complete amino acid sequence of pig kidney fructose-1,6-bisphosphatase.
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DOI：
10.1073/pnas.79.23.7161
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发表时间：
1982-12
影响因子：
11.1
通讯作者：
F. Marcus;I. Edelstein;I. Reardon;R. Heinrikson
中科院分区：
文献类型：
--
作者：
F. Marcus;I. Edelstein;I. Reardon;R. Heinrikson
确定了猪肾果糖-1,6-二磷酸酶（d -果糖-1,6-二磷酸1-磷酸水解酶，EC 3.1.3.11）亚基的共价结构。多肽链上335个氨基酸残基的位置主要是基于s -羧甲基化蛋白产生的8个纯化溴化氰片段的自动Edman降解。为了确定最大的溴化氰片段（154个残基）的氨基酸序列，需要对精氨酸残基的色氨酸切割和Asp-Pro肽键的温和酸切割获得的亚片段进行额外的分析。溴化氰片段的比对是通过内源性蛋白酶有限蛋白水解产物的分析和从S-[14C]羧甲基化果糖-1,6-二磷酸酶分离的色氨酸肽的表征来完成的。这一序列信息使鉴定猪肾果糖-1,6-双磷酸酶的几个功能和结构上有意义的活性位点成为可能。
The covalent structure of the pig kidney fructose-1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrolase, EC 3.1.3.11) subunit has been determined. Placement of the 335 amino acid residues in the polypeptide chain was based largely on automated Edman degradation of eight purified cyanogen bromide fragments generated from the S-carboxymethylated protein. The determination of the amino acid sequence of the largest cyanogen bromide fragment (154 residues) required additional analysis of subfragments obtained by tryptic cleavage at arginyl residues and by mild acid cleavage of an Asp-Pro peptide bond. Alignment of the cyanogen bromide fragments was accomplished by analysis of a product of limited proteolysis by an endogenous protease and by characterization of the tryptic peptides isolated from S-[14C]carboxymethylated fructose-1,6-bisphosphatase. This sequence information has permitted the identification of several reactive sites of functional and structural significance in pig kidney fructose-1,6-bisphosphatase.