STRUCTURES OF CATALYTIC DOMAINS OF BLOOD PROTEINS

血液蛋白催化域的结构

• 批准号: 2220938
• 负责人: ALEXANDER TULINSKY
• 金额: $ 17.78万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2220938
• 关键词: X ray crystallography; activation product; active sites; chemical association; chemical binding; coagulation factor IX; coagulation factor VII; coagulation factor X; conformation; crystallization; enzyme complex; enzyme inhibitors; enzyme structure; fibrinogen; heparin; hirudins; protein C; protein structure function; prothrombin; thrombin; thromboplastin

The general thrust of the long range plans is to pursue crystallographically structural aspects of molecules of blood coagulation and fibrinolysis with the aim of relating them to functionality at the molecular level. Our research program has been exclusively devoted to the area for about the past 15 years and has made significant progress by determining the structures of all but one of the autonomous domains of these molecules (gamma-carboxyglutamic, kringle, epidermal growth factor, catalytic), along with many informative derivative studies involving them or combinations thereof. We will now extend the work to include additional catalytically active multidomain structures, which will also address domain-domain associations in a more general way. Since these molecules interact with macromolecular substrates, inhibitors and cofactors, the manner of such interactions will be revealed through the structure determinations of molecular complexes, including rationally designed mimetic molecules. Specific studies targeted include: (l) the inactive precursor of alpha-thrombin (prethrombin 2), (2) Serl95Ala anhydro thrombin and hirugen-anhydro thrombin to capture the active site conformational change accompanying fibrinogen recognition exosite binding, (3) a conformationally restricted non-peptide mimetic based on the unusual N-terminal hirudin interaction in the active site of thrombin, (4) the structure determination of Factor Xa, (a) in the presence of Ca+2 ions and (b) in complex with the second Kunitz domain of tissue factor pathway inhibitor and (5) the structure determination of the potent Factor Xa inhibitor tick anticoagulant protein. Crystallization searches are also underway with Factors VIIa and IX and protein C and activated protein C to extend the scope of the work to other areas of the blood coagulation cascade. Diffraction quality single crystals of many of the above have been grown and their X-ray diffraction patterns examined in a preliminary way and three intensity data sets have been measured that suggest a high probability of success.

远程计划的一般性是追求 血液凝结分子的结构结构方面 和纤维蛋白溶解，目的是将它们与功能联系起来 分子水平。 我们的研究计划专门用于 大约过去15年的区域，并取得了重大进展 确定除一个自治领域之一之外的所有结构 这些分子（γ-羧基谷氨酸，kringle，表皮生长因子， 催化），以及许多涉及它们的衍生衍生研究 或组合。现在，我们将扩展工作以包括其他 催化活性多域结构，这也将解决 域状域的关联以更一般的方式。由于这些分子 与大分子底物，抑制剂和辅因子相互作用， 这种互动的方式将通过结构揭示 分子复合物的确定，包括合理设计的 模拟分子。针对性的具体研究包括：（l）无效 α-凝血酶的前体（凝血酶2），（2）Serl95ala anhydro 凝血酶和海龙 - 抗凝血酶以捕获活性部位 伴有纤维蛋白原识别外源结合的构象变化， （3）基于不寻常的构象受限的非肽模拟物 凝血酶活性部位的N末端Hirudin相互作用，（4） Xa因子的结构测定（a）在Ca+2离子存在下的结构确定和 （b）与第二个组织因子途径的第二个Kunitz结构域中 抑制剂和（5）有效因子XA的结构测定 抑制剂滴答抗凝蛋白。结晶搜索也是 VIIA和IX和蛋白C的因素正在进行中，并激活的蛋白C 将工作范围扩展到血液凝结的其他区域 级联。以上许多的衍射质量单晶具有 被种植，其X射线衍射模式在初步中检查 已经测量了方式和三个强度数据集，这表明很高 成功的概率。