Basic research for pathogenesis of type 1 von Willebrand disease

1型冯维勒布兰德病发病机制的基础研究

• 批准号: 12671011
• 负责人: MATSUI Taei
• 金额: $ 1.98万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671011/
• 关键词: von Willebrand disease; von Willebrand factor; ABO blood group; lectin; bitiscetin; platelet aggregation; Upshaw-Schulman syndrome; kaouthiagin; フォン・ビルブランド因子; Upshaw-Schulman症候群; VWF切断プロテアーゼ; ELISA; フォン・ビルブランド病; 一次構造; メタロプロテアーゼ; ジスインテグリン

We have screened lectins that can recognize the difference in the carbohydrate structures on von Willebrand factor (VWF) from normal individuals with each ABO blood group and patients with type 1 von Willebrand disease (VWD). Furthermore, we have studied the structure and function of snake venom proteins affecting human hemostasis by interacting with VWF and platelet.1) Among blood group-reconizing lectins examined, HPA and UEA-I were useful for detection and fractionation of blood group A and O(H) sugar chains on VWF, respectively (BBA, 1525: 50-57, 2001). We have found that the sialic acid-recognizing lectins (SSA and SNA) and the H structure-recognizing lectins (TJA-I and UEA-I) preferentially bound to VWF from group O and type 1 VWD plasmas, suggesting that these VWFs contain terminal sialic acid residues and the H structure in a higher amount.2) We have determined the amino acid sequence of kaouthiagin which specifically cleaves VWF, and found that its Cys-rich domain has the functional disintegrin-like activity (Biochemistry 40: 4503-4511, 2001). We have determined the crystal structure of bitiscetin which modulates VWF to induce platelet aggregation, and found that it has a central conecave structure rich in negatively charged residues, implying that it might be the VWF-binding site (Biochemistry 40: 13592-13597, 2001).3) We have found that the deficiency of physiological VWF-cleaving metalloproteinase in plasma causes the presence of ultra-high molecular mass multimers of VWF subunits resulting in the Upshaw-Schulman syndrome (USS), the congenital thrombotic thrombocytopenic purpura (TTP). (Int. J. Hematol, 74: 101-108, 2001, Int. J. Hematol., 75: 25-34, 2002).

我们已经筛选了凝集素，可以识别不同的碳水化合物结构的血管性血友病因子（VWF）从正常人与每个ABO血型和1型血管性血友病（VWD）患者。1）在所检测的血型识别凝集素中，HPA和UEA-I分别可用于检测和分离VWF上的血型A和O（H）糖链（BBA，1525：50-57，2001）。我们发现唾液酸识别凝集素（SSA和SNA）和H结构识别凝集素（TJA-I和UEA-I）优先结合来自O型和1型VWD血浆的VWF，表明这些VWF含有末端唾液酸残基和更高亲和力的H结构。2）我们已经确定了特异性切割VWF的Kaouthiagin的氨基酸序列，并发现其富含Cys的结构域具有功能性去整合素样活性（Biochemistry 40：4503-4511，2001）。我们测定了调节VWF诱导血小板聚集的bitiscycloid的晶体结构，发现它具有一个富含负电荷残基的中心锥状结构，暗示它可能是VWF的结合位点（生物化学40：13592-13597,2001）。3）我们已经发现，血浆中生理性VWF切割金属蛋白酶的缺乏导致超VWF亚基的高分子量多聚体导致Upshaw-Schulman综合征（USS）、先天性血栓性血小板减少性紫癜（TTP）。(Int. J. Hematol.，74：101-108，2001，Int. J. Hematol.，75：25-34，2002）。

项目成果

S.Kinoshita, et al.: "Upshaw-Schulman syndrome revisited : a congenital deficiency of von Willebrand factor-cleaving protease, and its correction with fresh frozen plasma"Int.J.Hematol.. 74. 101-108 (2001)

S.Kinoshita 等人：“Upshaw-Schulman 综合征重温：冯维勒布兰德因子裂解蛋白酶的先天性缺陷及其用新鲜冰冻血浆的校正”Int.J.Hematol.. 74. 101-108 (2001)

松井太衛(共著): "第7回血液アゴラ"メディカル・ジャーナル社. 85-91 (2000)

Tae Matsui（合著者）：“7th Blood Agora”Medical Journal Inc. 85-91 (2000)

H. Yagi, et al.: "Plasma of patients with Upshaw-Schulman syndrome enhances the aggregation of normal platelets under high shear stress."Br. J. Haematol.. 115. 991-997 (2001)

H. Yagi 等人：“Upshaw-Schulman 综合征患者的血浆在高剪切应力下增强了正常血小板的聚集。”Br。

Y. Sakurai, et al.: "Inhibition of human platelet aggregation by L-aminoacid oxidase purified from Naja naja kaouthia venom."Toxicon. 39. 1827-1833 (2001)

Y. Sakurai 等人：“从 Naja naja kaouthia 毒液中纯化的 L-氨基酸氧化酶抑制人血小板聚集。”Toxicon。

Kinoshita, S., et al.: "Upshaw-Schulman syndrome revisited : a congenital deficiency of von Willebrand factor-cleaving protease, and its correction with fresh frozen plasma"Int.J.Hematol.. 74(1). 101-108 (2001)

Kinoshita, S., 等人：“Upshaw-Schulman 综合征重温：冯维勒布兰德因子裂解蛋白酶先天性缺陷及其用新鲜冰冻血浆的校正”Int.J.Hematol.. 74(1)。