Studies on Molecular Abnormality of Blood Coagulation and Fibrinolytic Factors

凝血及纤溶因子分子异常研究

• 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

(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digest of the abnormal thrombin indicated that Arg-418 (equivalent to Asn-101 in the chymotrypsin numbering system) had been replaced by Trp.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)两例异常纤维蛋白原川口和大阪均为纤维蛋白原异常杂合子，分别命名为川口纤维蛋白原和大阪纤维蛋白原，均为纤维蛋白原异常杂合子。通过采用一种新的策略，利用来自Achromobacter lyticus M497-1的一种丝氨酸蛋白酶Lysyl endopeptidase，我们从S-吡啶乙基化的A&lt；α&gt；链的消化液中分离出异常的A&lt；α&gt；1-29肽，并确定在异常纤维蛋白原川口和大阪的A&lt；α&gt；Arg-16被半胱氨酸取代。(2)凝血酶原德岛：用色氨酸取代精氨酸-418，损害衍生的凝血酶托克岛的纤维蛋白原凝血活性；对一种遗传性异常的凝血酶原--凝血酶原德岛进行了结构研究，以确定其转化为凝血酶后纤维蛋白原凝血活性降低的原因。所使用的凝血酶原样本来自杂合子，但仅含有缺陷的凝血酶原分子，因为患者同时患有凝血酶原紊乱症和低凝血酶原血症。对异常凝血酶赖氨酰内肽酶的氨基酸序列分析表明，Arg-418(相当于胰凝乳酶编号系统中的ASN-101)已被Trp取代。这种氨基酸替代可以由Arg-418(CGG-&GT；TGG)密码子的单核苷酸变化引起。在凝血酶原德岛的凝血酶部分发现的Arg-&GT；Trp替代似乎减少了它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，并解释了先证者中观察到的反复出血事件。

项目成果

Morita, T., Mizuguchi, J., Kawabata, S., and Iwanaga, S.: "Proteolytic Cleavage of Vitamin K-dependent Bovine Plasma Protein S by <alpha> -Thrombin and Plasma Serine Protease." J. Biochem.99. 561-568 (1986)

Morita, T.、Mizuguchi, J.、Kawabata, S. 和 Iwanaga, S.：“<α>-凝血酶和血浆丝氨酸蛋白酶对维生素 K 依赖性牛血浆蛋白 S 进行蛋白水解切割”。

宮田敏行,岩永貞昭 著,小谷正雄,大井龍夫,次田晧,崎山文夫,岩永貞昭 編: "蛋白質・DNAのデータバンクと情報解析" 共立出版(東京), (1986)

Toshiyuki Miyata、Sadaaki Iwanaga、Masao Kotani、Tatsuo Oi、Akira Tsugita、Fumio Sakiyama、Sadaaki Iwanaga（编）：“蛋白质/DNA 数据库和信息分析”Kyoritsu Shuppan（东京），（1986）

Miyata, T., Matsumoto, H., Hattori, M., Sakaki, Y., and Iwanaga, S.: "Two Types of Coagulogen mRNAs Found in the Horseshoe Crab (Tachypleus tridentatus) Hemocytes: Its Molecular Coning and Nucleotide Sequence." J. Biochem.100. 213-220 (1986)

Miyata, T.、Matsumoto, H.、Hattori, M.、Sakaki, Y. 和 Iwanaga, S.：“鲎 (Tachypleus tridentatus) 血细胞中发现的两种凝固蛋白原 mRNA：其分子圆锥和核苷酸序列。

Morita,T.: J.Biochem.99. 561-568 (1986)

森田，T.：J.Biochem.99。

岩永貞昭 著,日比野進 監修: "血液学" 丸善(東京), (1985)

岩永贞明编剧，日比野进监督：《血液学》丸善（东京），（1985）