Studies on Molecular Abnormality of Blood Coagulation and Fibrinolytic Factors

凝血及纤溶因子分子异常研究

基本信息

批准号：
60480497
负责人：
IWANAGA Sadaaki
金额：
$ 4.35万
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 (B)
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/en/grant/KAKENHI-PROJECT-60480497/
关键词：
Fibrinogen Prothrombin Abnormal molecule Blood coagulation factor セリンプロテアーゼ前駆体

项目摘要

(1) DEFECTIVE RELEASE OF FIBRINOPEPTIDE A DUE TO SUBSTITUTION OF A <alpha> ARGININE-16 BY CYSTEINE IN TWO ABNORMAL FIBRINOGENS KAWAGUCHI AND OSAKA: The propositi of two abnormal fibrinogens, designated as fibrinogen Kawaguchi and fibrinogen Osaka, are both heterozygotes for the abnormality of fibrinogen characterized by defective release of fibrinopeptide A (FPA). By employing a new strategy of utilizing lysyl endopeptidase, a seryl protease derived from Achromobacter lyticus M497-1, we separated aberrant A <alpha> 1-29 peptides from the digests of the S-pyridylethylated A <alpha> chain and identified substituion of A <alpha> Arg-16 by Cys in both abnormal fibrinogens Kawaguchi and Osaka.(2) PROTHROMBIN TOKUSHIMA: A REPLACEMENT OF ARGININE-418 BY TRYPTOPHAN THAT IMPAIRS THE FIBRINOGEN CLOTTING ACTIVITY OF DERIVED THROMBIN TOKUSHIMA; Structural studies on a hereditarily abnormal prothrombin, prothrombin Tokushima, have been performed to identify the difference responsible for its reduced fibrinogen clotting activity upon conversion to thrombin. The prothrombin sample used was from a heterozygote but contained exclusively a defective prothrombin molecule, since the patient was heterozygous for both dysprothrombinemia and hypoprothrombinemia. Amino acid sequence analysis of a peptide isolated from a lysyl endopeptidase digest of the abnormal thrombin indicated that Arg-418 (equivalent to Asn-101 in the chymotrypsin numbering system) had been replaced by Trp. This amino acid substitution can result from a single nucleotide change in the codon for Arg-418 (CGG-> TGG). The Arg-> Trp replacement found in the thrombin portion of prothrombin Tokushima appears to reduce its interaction with various substrates including fibrinogen and platelet receptors, and accounts for the recurrent bleeding episode observed in the propositus.

（1）在两个异常纤维纤维菌中用半胱氨酸取代<Alpha>精氨酸16，纤维蛋白肽A的缺陷释放：川口和大阪：两种异常纤维蛋白的提议，指定为As fibrinegen kawaguchi和Fibibrinsigent byokgen byogenoration frotheogentator fortheogotient fortheozygotient fortheozygotient fortheogote fortheozygotipthe osakygotient纤维蛋白肽A（FPA）的有缺陷释放。通过采用一种利用蛋白质内肽酶的新策略，该蛋白酶是源自achromobacter liticus m497-1的塞基蛋白酶，我们将异常的A <alpha> 1-29肽与S-吡啶基乙基乙基乙基的消化物的消化中的<alpha>的消化作用分开（2）凝血酶原tokushima：色氨酸替代精氨酸418，这会损害衍生的凝血酶tokushima的纤维蛋白原凝结活性；已经对遗传异常凝血酶原凝血酶原Tokushima进行了结构研究，以确定导致其转化为凝血酶后纤维蛋白原凝血活性减少的差异。使用的凝血酶原样品来自杂合子，但仅包含缺陷的凝血酶原分子，因为患者的染色体蛋白酶血症和低蛋白酶血症都是杂合的。从异常凝血酶的赖氨酸内肽酶消化物中分离出的肽序列分析表明，agn-418（相当于chymotrypsin编号系统中的ASN-101）已被TRP替换。该氨基酸取代可能是由ARG-418（CGG-> TGG）的密码子中的单个核苷酸变化引起的。在凝血酶原Tokushima的凝血酶凝血酶部分中发现的Arg-> TRP替换似乎减少了其与包括纤维蛋白原和血小板受体在内的各种底物的相互作用，并解释了在propositus中观察到的反复出血性发作。