Research for the primary structures of human vitamin D-binding proteins.

研究人类维生素 D 结合蛋白的一级结构。

• 批准号: 60440042
• 负责人: MATSUMOTO Hideo
• 金额: $ 0.51万
• 依托单位: Osaka Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60440042/
• 关键词: Gc; Vitamin D-binding protein; Serum protein; Primary structure; Subtype; Amino acid substitution; 多型現象; ビタミンD結合; Gc蛋白; 遺伝標識

Gc protein is one of human serum protein, which is widely available in the field of legal medicine and human genetics, and has several phenotypes controlled by three codominat genes (Gc2, Gc1F, and Gc1S) on isoelctric focusing electrophoresis. This protein, which is called vitamin D-binding protein (DBP) by the character, has important functions under the affinity of actin in serum or on lymphocyte. The primary structure of DBP, which many researchers in the world could not clarify in spite of many approaches, was determined through the base sequence analyses of that cDNA by Yang on 1985. We intended to confirm this amino acid sequence speculated by the base sequence and to elucidate the amino acid substitutions between Gc subtypes. DBPs were purified from human sera by salting and chromatography. The BrCN peptides of carboxymethylated DBP were separated by hiph-performance liquid chromatography, and further purification of the enzymatic digests of BrCN peptide was carried out on reversed-phase chromatography. We confirmed the primary structure of DBP (Gc2) by the amino acid composition analyses and the amino acid sequence analyses of these peptides. The sequence analysis of DBP carrying Gc1F or 1S was done by the same procedures. From these results, the amino acid substitutions between DBP subtypes are GLY-GLU-GlU (Gc2-1F-1S) at position 152, ASP-ASP-GLU at position 416, and LYS-THR-THR at position 420.

Gc蛋白是人血清蛋白的一种，广泛应用于法律医学和人类遗传学领域，在等电聚焦电泳上具有由Gc2、Gc1F和Gc1S三个共显性基因控制的多种表型。该蛋白在肌动蛋白的作用下或在淋巴细胞上具有重要的作用，其性质称为维生素d结合蛋白（DBP）。DBP的一级结构是由Yang于1985年通过对该cDNA的碱基序列分析确定的，目前世界上许多研究者通过许多方法都不能明确DBP的一级结构。我们打算通过碱基序列来证实这个氨基酸序列，并阐明Gc亚型之间的氨基酸替换。采用盐法和色谱法从人血清中纯化DBPs。采用高效液相色谱法分离羧甲基化DBP的BrCN肽段，反相色谱法进一步纯化酶解的BrCN肽段。我们通过氨基酸组成分析和氨基酸序列分析确定了DBP （Gc2）的一级结构。携带Gc1F或1S的DBP序列分析方法相同。从这些结果来看，DBP亚型之间的氨基酸替换为152位的GLY-GLU-GlU (Gc2-1F-1S)， 416位的ASP-ASP-GLU和420位的LYS-THR-THR。