Basic research for pathogenesis of type 1 von Willebrand disease

我们从不同ABO血型的正常人和1型血管性血友病（VWD）患者中筛选了能够识别血管性血友病因子（VWF）碳水化合物结构差异的凝集素。此外，我们还研究了蛇毒蛋白通过与VWF和血小板相互作用影响人止血的结构和功能。1)在检测的血型识别凝集素中，HPA和UEA-I分别可用于VWF上的A型和O(H)型糖链的检测和分离（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）优先与来自O组和1型VWD血浆的VWF结合，这表明这些VWF含有更多的末端唾液酸残基和H结构。2)我们测定了特异裂解VWF的kaouthiagin的氨基酸序列，发现其富含cys的结构域具有功能性崩解素样活性（Biochemistry 40: 4503- 4511,2001）。我们已经确定了通过调节VWF诱导血小板聚集的bitscittin的晶体结构，发现它具有一个富含负电荷残基的中心凹结构，这意味着它可能是VWF的结合位点（Biochemistry 40: 13592-13597, 2001）。3)我们发现血浆中生理性VWF切割金属蛋白酶的缺乏会引起VWF亚基的超高分子质量多聚体的存在，从而导致先天性血栓性血小板减少性紫癜（TTP）。(Int。中华血液学杂志，21(4):391 - 398,2001。j .内科杂志。科学，75:25-34,2002)。

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).