Molecular basis for the fibrinogen structure and functions-Analysis Of hereditary dysfibrinogens and their application to the study

纤维蛋白原结构和功能的分子基础-遗传性异常纤维蛋白原的分析及其在研究中的应用

• 批准号: 11694308
• 负责人: MATSUDA Michio
• 金额: $ 2.11万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694308/
• 关键词: fibrinogen; hereditary dysfibrinogen; bleeding tendency; fibrinogen structure; extra carbohydrate; fibrin nets; fibrin ultra-structure; gene analysis; 余剩糖鎖; 遺伝性異常フィブリノゲン; フィブリノゲン異常症; 血栓性素因; 糖付加構造; 電顕学的分析; フィブリンの超微細構造; 停止コドンの消失

1. The Molecular Mechanisms of Extra Asn-linked OligosaccharidesBy transmission electron microscopy (TEM), we have studied the role of extra sugar moiteis linked to Asn residues due to creation of an Asn-X-Thr/Ser type sequence owing to an amino acid substitutionIn fibrinogen Asahi, the extra sugar moities linked to γAsn-308 residues due to a γMet-310 to Thr substitution are suggested to interfere with the E-D binding and cross-linking of the fibrin γ-chain. Indeed, removal of the sugar moities resulted in the formation of well ordered fibrin networks with appropriate branching, although the D : D association still remained owing to the point mutation. Thus, we conclude that the extra sugar moieties are largely responsible for the functional abnormality in this dysfibrinogen.On the contrary, a high proportion of disialylated oligosaccharide in fibrinogen Lima (78%) generates electric repulsive forces due to sialic acids and disturbs lateral association of fibrin protofibrils on polymer … More ization. Removal of sialic acids alone lead to formation normal fibrin ultra structures as has been suggested by biochemical studies.2. Characterization of a unique dysfibrinogen, fibrinogen Osaka VI with a 12 residue extension of the Bb-chain.In a dysfibrinogen derived from a 36-year-old woman who had suffered from severe bleeding on two occasions of childbirth, we have identified a 12-residue extension of the Bβ-chain due to a transition of (TAG) for stop to (AAG) for Lys. Thus, additional 36 base pairs are translated. Interestingly, one or two disulfide bridges are formed between two corresponding Cys residues at the second position from the new C-terminus, leading to formation of end-linked fibrinogen homodimers, which are aligned either in parallel or in tandem as demonstrated by TEM.Inclusion of these fibrinogen dimers in protofibrils seems to be related to the formation of highly branched and fragile fibrin clots.The results have been reported in Blood 96(12) : 3779-3785, 2000 and also in the Annals of New York Academy of Sciences, in press. Less

在纤维蛋白原朝日中，由于γMet-310到Thr的取代而与γAsn-308残基相连的额外糖基被认为干扰了纤维蛋白γ链的E-D结合和交联性。的确，糖基团的去除导致了具有适当分支的有序的纤维蛋白网络的形成，尽管由于点突变，D：D关联仍然存在。因此，我们得出结论，额外的糖部分是导致这种纤维蛋白原功能异常的主要原因。相反，纤维蛋白原LIMA中高比例的二唾液酸寡糖(78%)由于唾液酸而产生电排斥力，并干扰纤维蛋白原与聚合物…的侧向结合更加本土化。生物化学研究表明，单独去除唾液酸可形成正常的纤维蛋白超微结构。一种独特的纤维蛋白原，大阪六号纤维蛋白原，具有12个残基的BB链延伸。在一个来自一位36岁妇女的纤维蛋白原中，我们发现了Bβ链的12个残基的延伸，这是由于(TAG)的停止转变为(AAG)的赖氨酸。因此，额外的36个碱基对被翻译。有趣的是，在距离新的C末端第二位的两个相应的Cys残基之间形成了一个或两个二硫键，导致末端连接的纤维蛋白原同源二聚体的形成，如TEM所证明的那样，这些纤维蛋白原二聚体平行或串联排列。这些纤维蛋白原二聚体似乎与高度支化和脆弱的纤维蛋白凝块的形成有关。这一结果已在血液96(12)：3779-3785,2000和纽约科学院年鉴中报道。较少

项目成果

Kant M Matsuda: "A novel strategy for the tumor angiogenesis-targeted gene therapy : Generation of angiostatin from endogenous plasminogen by protease gene transfer"Cancer Gene Therapy. (in press).

Kant M Matsuda：“肿瘤血管生成靶向基因治疗的新策略：通过蛋白酶基因转移从内源性纤溶酶原生成血管抑制素”癌症基因治疗。

Isao Kohno: "A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin : Its application to the measurement of granulocyte-derived elastase-digests in plasma"Blood. 95. (2000)

Isao Kohno：“一种对人纤维蛋白原和纤维蛋白的粒细胞来源的弹性蛋白酶片段 D 种具有特异性的单克隆抗体：其在测量血浆中粒细胞来源的弹性蛋白酶消化物中的应用”血液。

Matsuda M: "Structure and function of fibrinogen inferred from hereditary dysfibrinogens."Fibrinolysis & Proteolysis. 14. 436-447 (2000)

松田 M：“从遗传性纤维蛋白原异常推断纤维蛋白原的结构和功能。”纤维蛋白溶解

Mimuro J, Kawata Y, Niwa K, Muramatsu S, Madoiwa S, Takano H, Sugo T, Sakata Y, Sugimoto T, Nose K, Matsuda M: "A new type of Ser substitution for γ Arg-275 in fibrinogen Kamogawa I characterized by impaired fibrin assembly."Thromb Haemost. 81. 940-944 (1

Mimuro J、Kawata Y、Niwa K、Muramatsu S、Madoiwa S、Takano H、Sugo T、Sakata Y、Sugimoto T、Nose K、Matsuda M：“纤维蛋白原 Kamokawa I 中 γ Arg-275 的新型 Ser 替代物的特征纤维蛋白组装受损。“血栓 Haemost。81. 940-944 (1

Kohno I, Inuzuka K, Itoh Y, Nakahara K, Eguchi Y, Sugo T, Soe G, Sakata Y, Murayama H, Matsuda M: "A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin : Its application to the measurement of

Kohno I、Inuzuka K、Itoh Y、Nakahara K、Eguchi Y、Sugo T、Soe G、Sakata Y、Murayama H、Matsuda M：“一种对人纤维蛋白原和纤维蛋白的粒细胞来源的弹性蛋白酶片段 D 种具有特异性的单克隆抗体