权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Structural studies on Functional Abnormality of Factor XII and Fibrinogen.

因子 XII 和纤维蛋白原功能异常的结构研究。

基本信息

批准号：
62580126
负责人：
ITO Akio
金额：
$ 0.96万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

(1) Fibrinogen Nagoya, a Replacement of Glutamine-329 by Arginine in the -chain. That Impairs the Polymerization of Fibrin Monomer: Structural studies on a hereditary abnormal fibrinogen, fibrinogen Nagoya were performed to identify the abnormality responsible for the impaired polymerization of fibrin nomomer. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. fibrinogen Nagoya showed the presence of an extra protein band with an apparent molecular weight of 49,500 in addition to the normal three subunit chains. Amino acid sequence analysis of a peptide isolated from a lysyl endopeptidase digest of one of the cnbr fragments derived from fibrinogen nagoya indicated that Gln-329 in the -chain had been replaced by Arg. This substitution can be explained by a single uncleotide chainge in the dodon for Gln-329 (CAG - CGG). We conclude that GLN-329 in the -chain is indispensable for the normal polymerization of fibrin monomer.(2) Blood Clontting Factor IX Niigata: Substitution of Alanine-390 by Valine in the Catalytic Domain: Factor IX Niigata is s mutant factor IX responsible for the moderately severe hemophilia B in a patient who has a normal level of factor IX antigen with reduced clotting activity (1-4% of normal). We reported previously that the purified mutant protein could be converted to the factor IXa form by factor XIa/Ca^<2+> at a rate similar to that in the case of normal factor IX, but the resulting mutant factor IXa could not activate factor X in the presence of factor XIII, Ca^<2+>, and phospholipids. In the present study, we analyzed factor IX Niigata at the structural level to elucidate the molecular abnormality responsible for the loss of clotting activity. Amino acid sequence analysis of a peptide obtained on lysyl endopeptidase digestion, coupled with subsequent SP-V8 digestion, demonstrated that the alanine at position 390 was substituted by valine in the catalytic domain of the factor IX Niigata molecule.

(1)纤维蛋白原名古屋，-链中谷氨酰胺-329被精氨酸取代。损害纤维蛋白原的聚合：对遗传性异常纤维蛋白原名古屋进行结构研究，以确定导致纤维蛋白原聚体聚合受损的异常。还原条件下十二烷基硫酸钠-聚丙烯酰胺凝胶电泳。纤维蛋白原名古屋显示除了正常的三个亚基链之外，还存在表观分子量为49，500的额外蛋白质条带。从来自纤维蛋白原名古屋的cnbr片段之一的赖氨酰内肽酶消化物中分离的肽的氨基酸序列分析表明，在-链中的Gln-329已被Arg取代。这种取代可以用Gln-329（CAG-CGG）的dodon中的单个核苷酸链来解释。我们的结论是，GLN-329的-链是不可缺少的纤维蛋白单体的正常聚合。(2)血液克隆因子IX新泻：在催化结构域中丙氨酸-390被缬氨酸取代：因子IX新泻是突变的因子IX，其导致患者中的中度重度血友病B，该患者具有正常水平的因子IX抗原，具有降低的凝血活性（正常的1 - 4%）。我们以前曾报道过，纯化的突变蛋白质可以通过因子XIa/Ca ^<2 +>以与正常因子IX相似的速率转化为因子IXa形式，但是在因子XIII、Ca ^<2 +>和磷脂存在的情况下，所得的突变因子IXa不能激活因子X。在本研究中，我们分析了第九因子新泻在结构水平上阐明的凝血活性的损失负责的分子异常。对赖氨酰内肽酶消化，再加上随后的SP-V8消化获得的肽的氨基酸序列分析表明，在因子IX新泻分子的催化结构域中，390位的丙氨酸被缬氨酸取代。