Experimental Studies on Fibrin Polymerization and Intrinsic Fibrinolysis

纤维蛋白聚合和内在纤维蛋白溶解的实验研究

• 批准号: 06671216
• 负责人: TANABE Gen
• 金额: $ 1.34万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06671216/
• 关键词: Fibrinogen; Fibrin polymerization; Fibrinopeptide B; 合成ペプチド; フィブリン

It is essential for blood clot generation that fibrinogen is converted into fibrin as a result of thrombin cleaving peptides from the amino-terminal ends of the alpha and beta chains. However, the details required for a full mechanistic explanation of the fibrin formation including the polymerization sites has not been clarified until now.In this project, we investigated the characterization of the fibrin polymerization site in the amino-terminus of the beta chain utilizing synthetic peptides. The binding of the synthetic peptide, Gly-His-Arg-Pro-amide, which corresponds to the amino-terminus of the beta chain, to fragment D1 was found to depend on the concentration of calcium ions, being maximal at 10^<-4>M and decreasing on either side of this at room temperature. In contrast, the effect of calcium ions on the binding was different when examined using a fragment D1 sepharose column treated with sialidase. These results emphasize the close proximity of the calcium-binding and peptide-binding sites in the calboxy-terminus of beta chain to which the carbohydrate attaches. Furthermore, we found the synthetic peptide Gly-His-Arg to have a slight protective effect on the gamma chain remnant of fragment D1 against plasmin in the same fashion as calcium ions.These date indicate that one of the binding sites of the amino-terminal ends of beta chain is located in the carboxy-terminus of beta chain and the other is located in the carboxy-terminus of gamma chain.

由于凝血酶从 α 链和 β 链的氨基末端裂解肽，纤维蛋白原转化为纤维蛋白对于血凝块的生成至关重要。然而，迄今为止，对纤维蛋白形成的完整机制解释所需的细节（包括聚合位点）尚未阐明。在本项目中，我们利用合成肽研究了 β 链氨基末端纤维蛋白聚合位点的表征。发现合成肽Gly-His-Arg-Pro-amide(其对应于β链的氨基末端)与片段D1的结合取决于钙离子的浓度，在10^-4M处最大，并且在室温下在该浓度的两侧减少。相反，当使用用唾液酸酶处理的片段 D1 琼脂糖柱检查时，钙离子对结合的影响是不同的。这些结果强调了碳水化合物所附着的β链羧基末端的钙结合位点和肽结合位点非常接近。此外，我们发现合成肽Gly-His-Arg以与钙离子相同的方式对片段D1的γ链残余物对纤溶酶具有轻微的保护作用。这些数据表明β链氨基末端的结合位点之一位于β链的羧基末端，另一个位于γ链的羧基末端。

项目成果

山角健介: "フィブリノゲン異常症" 血液・腫瘍科. 32. 20-26 (1996)

Kensuke Yamazumi：“纤维蛋白原功能障碍”血液学和肿瘤学系 32. 20-26 (1996)。

山角健介: "Bβ-フラグメント" 臨床検査. 40・11. 170-171 (1996)

Kensuke Yamazumi：“Bβ-片段”临床测试40・11（1996）。

山角健介: "D-ダイマー" 血液・腫瘍科. 28. 285-289 (1994)

Kensuke Yamazumi：“D-二聚体”血液学和肿瘤学系 28. 285-289 (1994)。

山角健介: "血管壁への白血球粘着とフィブリノゲン" 血液・腫瘍科. 31. 147-150 (1995)

Kensuke Yamazumi：“白细胞粘附到血管壁和纤维蛋白原”血液学和肿瘤学系 31. 147-150 (1995)。

Kensuke, Yamazumi: "Leukocyte adhesion to vascular endothelium and fibtinogen." Hematology and Oncology (Tokyo). 31. 147-150 (1995)

Kensuke, Yamazumi：“白细胞粘附到血管内皮和纤维蛋白原。”