Analysis of plasmin generation and clot lysis of plasma fibrinogen purified from a heterozygous dysfibrinogenemia, BβGly15Cys (Hamamatsu II)
Analysis of plasmin generation and clot lysis of plasma fibrinogen purified from a heterozygous dysfibrinogenemia, BβGly15Cys (Hamamatsu II)
复制标题
DOI：
10.1097/mbc.0b013e328332aa09
复制
发表时间：
2009-12
影响因子：
1.1
通讯作者：
Yuka Kamijyo;M. Hirota-Kawadobora;K. Yamauchi;F. Terasawa;T. Honda;M. Ikeya;N. Okumura
中科院分区：
文献类型：
--
作者：
Yuka Kamijyo;M. Hirota-Kawadobora;K. Yamauchi;F. Terasawa;T. Honda;M. Ikeya;N. Okumura
我们发现了一种由Bβ Gly 15 Cys取代引起的杂合子异常纤维蛋白原血症，并将其命名为纤维蛋白原Hamamatsu II（H-II）。虽然先证者患有延髓梗死，但未发现其他血栓形成危险因素、反常脑梗死和动脉夹层，为了确定Bβ Gly 15 Cys替代是否延迟纤维蛋白凝块溶解，我们检测了先证者纤维蛋白原的凝块溶解和纤溶酶生成。通过免疫亲和层析从先证者和正常对照血浆中纯化纤维蛋白原，并用于以下实验：十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、纤维蛋白聚合、纤维蛋白凝块和纤维的扫描电镜观察、凝块溶解和组织型纤溶酶原激活剂介导的纤溶酶原激活。H-II血浆纤维蛋白原显示存在白蛋白结合变体形式，变体纤维蛋白原的二聚体分子，以及纤维蛋白聚合过程中侧向聚集的损伤。H-II纤维蛋白凝块显示出较低的密度的束和较薄的直径的纤维比在正常的纤维蛋白凝块。在覆盖纤溶酶的凝块溶解实验中，H-II纤维蛋白显示出与正常对照相似的溶解时间和溶解速率。此外，从凝血酶、组织型纤溶酶原激活物、纤溶酶原和H-II纤维蛋白原的混合物产生的纤溶酶也显示出与正常纤维蛋白原相似的速率。虽然先证者患有梗死，但本研究未观察到延迟性血凝块溶解，即血凝块对纤溶酶降解无抵抗力。因此，我们没有阐明Bβ Gly 15 Cys纤维蛋白原异常血症与动脉血栓形成之间的联系。
We found a heterozygous dysfibrinogenemia caused by the substitution of BβGly15Cys and designated it fibrinogen Hamamatsu II (H-II). Although the propositus suffered an infarction of the medulla oblongata, other thrombotic risk factors, paradoxical cerebral infarction, and arterial dissection were not found. To determine whether the delayed lysis of fibrin clots or not in the context of the BβGly15Cys substitution, we examined the clot lysis and plasmin generation of propositus' fibrinogen. Fibrinogen was purified from the propositus' and normal control plasma by immunoaffinity chromatography and was used for the following experiments: sodium dodecyl sulfate-polyacrylamide gel electrophoresis, fibrin polymerization, scanning electron microscopic observation of fibrin clot and fibers, clot lysis, and tissue-type plasminogen activator-mediated plasminogen activation. The H-II plasma fibrinogen showed the presence of albumin-binding variant forms, a dimeric molecule of variant fibrinogen, and impairment of lateral aggregation during fibrin polymerization. The H-II fibrin clot showed lower density of bundles and thinner diameters of fibers than in the normal fibrin clot. In the clot lysis experiments with overlaid plasmin, H-II fibrin showed a similar lysis period and lysis rate to the normal control. Moreover, plasmin generation from a mixture of thrombin, tissue-type plasminogen activator, plasminogen, and H-II fibrinogen also showed a similar rate to normal fibrinogen. Although the propositus suffered an infarction, the present study did not observe delayed clot lysis, that is, the clot was not resistant to plasmin degradation. Therefore, we did not clarify an association between the BβGly15Cys dysfibrinogenemia and arterial thrombosis.