STRUCTURES OF CATALYTIC DOMAINS OF BLOOD PROTEINS

血液蛋白催化域的结构

• 批准号: 2721498
• 负责人: ALEXANDER TULINSKY
• 金额: $ 3.06万
• 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/2721498
• 关键词: STRUCTURES; CATALYTIC; DOMAINS; BLOOD; PROTEINS

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.

长期计划的主旨是追求 凝血分子的晶体学结构 和纤维蛋白溶解，目的是将它们与 分子水平。 我们的研究项目一直致力于 在过去的15年里，该领域取得了重大进展， 确定除了一个自治域之外的所有自治域的结构， 这些分子（γ-羧基谷氨酸，kringle，表皮生长因子， 催化），沿着许多涉及它们的信息丰富的衍生研究 或其组合我们现在将扩大工作范围， 催化活性的多域结构，这也将解决 以更通用的方式进行域与域的关联。因为这些分子 与大分子底物、抑制剂和辅因子相互作用， 这种相互作用的方式将通过结构来揭示， 分子复合物的测定，包括合理设计的 模拟分子针对的具体研究包括：（l）非活性 α-凝血酶前体（凝血酶前体2），（2）Ser 195 Ala脱水物 凝血酶和水蛭素-脱水凝血酶来捕获活性位点 伴随纤维蛋白原识别外部位点结合的构象变化， (3)一种构象受限的非肽模拟物， 凝血酶活性部位N-末端水蛭素相互作用; 因子Xa的结构测定，（a）在Ca+2离子存在下， (b)与组织因子途径的第二Kunitz结构域复合 抑制剂和（5）有效因子Xa的结构测定 抑制剂蜱抗凝蛋白。结晶搜索也是 因子VIIa和IX以及蛋白C和活化蛋白C， 将工作范围扩展到血液凝固的其他领域 级联。衍射质量单晶的许多上述具有 生长和他们的X-射线衍射图案进行了初步检查， 方法和三个强度数据集已经测量，表明高 成功的可能性。

项目成果

The structure of a designed peptidomimetic inhibitor complex of alpha-thrombin.

设计的α-凝血酶拟肽抑制剂复合物的结构。

• DOI: 10.1093/protein/6.5.471
• 发表时间: 1993
• 期刊: Protein engineering
• 作者: Wu,TP;Yee,V;Tulinsky,A;Chrusciel,RA;Nakanishi,H;Shen,R;Priebe,C;Kahn,M
• 通讯作者: Kahn,M

Structure of a nonadecapeptide of the fifth EGF domain of thrombomodulin complexed with thrombin.

与凝血酶复合的血栓调节蛋白第五个 EGF 结构域的九肽结构。

• DOI: 10.1021/bi00250a006
• 发表时间: 1994
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Mathews,II;Padmanabhan,KP;Tulinksy,A;Sadler,JE
• 通讯作者: Sadler,JE

Synthesis, structure, and structure-activity relationships of divalent thrombin inhibitors containing an alpha-keto-amide transition-state mimetic.

含有α-酮酰胺过渡态模拟物的二价凝血酶抑制剂的合成、结构和构效关系。

• DOI: 10.1002/pro.5560050303
• 发表时间: 1996
• 期刊: Protein science : a publication of the Protein Society.
• 作者: Krishnan,R;Tulinsky,A;Vlasuk,GP;Pearson,D;Vallar,P;Bergum,P;Brunck,TK;Ripka,WC
• 通讯作者: Ripka,WC

Structures of thrombin retro-inhibited with SEL2711 and SEL2770 as they relate to factor Xa binding.

凝血酶的结构受到 SEL2711 和 SEL2770 的逆向抑制，因为它们与 Xa 因子结合有关。

• DOI: 10.1107/s0907444999000359
• 发表时间: 1999
• 期刊: Acta crystallographica. Section D, Biological crystallography
• 作者: Mochalkin,I;Tulinsky,A
• 通讯作者: Tulinsky,A

Crystallographic structure of a peptidyl keto acid inhibitor and human alpha-thrombin.

肽基酮酸抑制剂和人α-凝血酶的晶体结构。

• DOI: 10.1016/0968-0896(95)00096-y
• 发表时间: 1995
• 期刊: Bioorganic & medicinal chemistry
• 影响因子: 3.5
• 作者: Håkansson,K;Tulinsky,A;Abelman,MM;Miller,TA;Vlasuk,GP;Bergum,PW;Lim-Wilby,MS;Brunck,TK
• 通讯作者: Brunck,TK