ANTIADHESIVE AND ANTICOAGULANT ACTIVITY OF KININOGENS

HIGH(HK)和LOW的生物化学、细胞和分子生物学 (LK)将研究分子量激肽原以了解 结构-功能相互关联，因为它们与结合和作用有关 血液和血管细胞。D5已被确定为 与阴离子表面相互作用，并作为HKA与 中性粒细胞。缺失突变体以及合成的多肽应该 允许映射与中性粒细胞结合所需的序列(S) 和内皮细胞。含锌和不含锌的D5的物理研究 将执行加号。D6中的31个氨基酸序列包含 足够的信息来绑定到上的两个不连续的站点 前激肽释放酶。利用D6的多肽和缺失突变， 结合的最小序列和HK结合到的拓扑 前激肽释放酶将被确定。我们用这些多肽和同源词 前激肽释放酶抑制内皮细胞纤溶活性的研究 细胞。D3此前已被确定为一个细胞结合区 血小板、内皮细胞和中性粒细胞。外显子7、8和9编码 因为D3将被表达来检验假设其中一个外显子 Products包含有关中性粒细胞在 重链，而另一个负责抑制结合 凝血酶对血小板和内皮细胞的作用。综合， 构象受限的多肽将用于精细定位 基于D3分子模型的表面可达区域 香港。香港与Mac-1上的中性粒细胞结合，我们将进一步绘制 配体和受体，以更好地定义相互作用。我们最近做了 显示纤维蛋白原不与HK结合以竞争 内皮细胞，而抗α-NU-β3抗体不抑制 HK结合，排除了α-NU-β3作为HK的受体。 然而，我们发现Vitronectin和可溶性尿激酶受体 抑制HK结合，提示UK受体是结合部位 为了香港。我们将通过绘制香港地图来进一步检验这一假设 UK受体上的结合位点，并将HK上的结合位点映射到 受体。香港阻断凝血酶结合的机制 通过与凝血酶受体或GPIB相互作用的血小板 探索过了。激肽原缺乏的大鼠将被用来验证香港 或LK是一种重要的抗血栓蛋白。多肽来自 激肽原结构域将在大鼠身上测试其抗粘附性和 抗血小板的潜力。这种多肽可以作为模板，用于 类似肽的化合物，这应该是治疗脓毒症的， 关节炎、遗传性血管水肿和牙周病 溶栓治疗后再闭塞。

The biochemistry, cell and molecular biology of high (HK) and low (LK) molecular weight kininogens will be studied to learn about structure-function correlates as they relate to binding to and effects on blood and vascular cells. D5 has been identified as the region that interacts with anionic surfaces and as a binding site for HKA to neutrophils. Deletion mutants as well as synthesized peptides should allow mapping of the sequence(s) required for binding to neutrophils and endothelial cells. Physical studies of D5 with and without Zn plus plus will be performed. A 31 amino acid sequence in D6 contains sufficient information to bind to two noncontinuous sites on prekallikrein. Using peptides and deletion mutagenesis of D6, the minimal sequences for binding and the topology of HK binding to prekallikrein will be determined. We use these peptides and cognate peptides of prekallikrein to down-regulate fibrinolysis on endothelial cells. D3 has previously been identified as one cell binding region for platelets, endothelial cells and neutrophils. Exons 7, 8, and 9 coding for D3 will be expressed to test the hypothesis that one of the exon products contains all of the information for neutrophil binding on the heavy chain, while another is responsible for inhibiting the binding of thrombin to platelets and endothelial cells. Synthesized, conformationally restrained peptides will be used for fine mapping based on surface-accessible regions of a molecular model of D3 of HK. HK binds to neutrophils on Mac-1, and we will further map the ligand and receptor to better define the interaction. We have recently shown that fibrinogen does not compete with HK binding to endothelial cells, and an antibody to alpha nu beta 3 does not inhibit HK binding, ruling out alpha nu beta 3 as the receptor for HK. However, we found that vitronectin and soluble urokinase receptor inhibit HK binding, suggesting that the UK receptor is the binding site for HK. We will further test this hypothesis by mapping the HK binding site on the UK receptor and map the binding site on HK to the receptor. The mechanism by which HK blocks thrombin binding to platelets by interaction with the thrombin receptor or GPIb will be explored. Kininogen-deficient rats will be used to verify whether HK or LK is an important anti-thrombotic protein. Peptides from kininogen domains will be tested in rats for their antiadhesive and antiplatelet potential. Such polypeptides could serve as templates for peptidomimetic compounds, which should be therapeutic in sepsis, arthritis, hereditary angioedema, and gingival disease in addition to reocclusion after thrombolytic therapy.