MACROMOLECULAR DOMAIN INTERACTIONS OF BLOOD PROTEINS

我们长期计划的主旨是追求 凝血分子的结晶学结构 和纤溶作用，目的是将它们与 分子水平。我们的研究计划目前专门致力于 这片区域。我们已经取得重大进展，通过确定 除一个以外的所有这些重要的自治域的结构 分子。由于血液蛋白质通常是多结构域结构，我们 现在将解决域-域交互的问题。此外, 因为它们与其他大分子底物、抑制剂、 辅因，...，我们将揭示其中一些相互作用的方式 通过分子络合物的结构测定。具体的 要研究的域-域相互作用是：(L)kringle-kringle (纤溶酶原(PG)Kringles 4-5，PG Kringles 1-2-3)，(2)Kringle催化 结构域(凝血酶前1，凝血酶原2-凝血酶原2)，(3) 表皮生长因子样(EGF)-Kringle相互作用(EGF-尿激酶 Kringle)，(4)EGF-EGF和EGF-催化域(血栓调节蛋白(TM)EGF 4- 5-6-凝血酶，TM(408-426)环-凝血酶)。大分子相互作用 含碳水化合物将通过低分子肝素测定 凝血酶与尿激酶环的络合物及新的 已发现的纳米分子单链DNA-凝血酶相互作用 通过15聚体DNA-凝血酶复合体的结构揭示。我们还将 完成金属离子与GLA(伽马峰)结合的研究 (羧基谷氨酸)磷脂结合区与血液分子结合 PT、MG-和TB-片段的结构测定。这两个 分子表现出与钙碎片1相似的假想性质。然而， 我们过去一段时间的工作表明，这三个人很可能 不同的三维结构。衍射光质量单一 上述许多晶体已经被生长出来，它们的X射线 初步研究了衍射图。

The general thrust of our 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 is presently exclusively devoted to the area. We have already made significant progress by determining the structures of all but one of the important autonomous domains of these molecules. Since blood proteins are generally multi-domain structures, we will now address the problem of domain-domain interactions. In addition, since they interact with other macromolecular substrates, inhibitors, cofactors,..., we will reveal the manner of some of these interactions through the structure determinations of molecular complexes. The specific domain-domain interactions to be investigated are: (l) kringle-kringle (plasminogen (PG) kringles 4-5, PG kringles 1-2-3), (2) kringle-catalytic domain (prethrombin1, prothrombin (PT) kringle2-prethrombin2), (3) epidermal growth factor-like (EGF)-kringle interactions (EGF-urokinase kringle), (4) EGF-EGF and EGF-catalytic domain (thrombomodulin (TM) EGF 4- 5-6-thrombin, TM (408-426) loop-thrombin). Macromolecular interactions with carbohydrates will be determined through low molecular weight heparin complexes with thrombin and the kringle of urokinase and the newly discovered nanomolar single stranded DNA-thrombin interaction will be revealed by the structure of a 15 mer DNA-thrombin complex. We will also complete our studies of metal ion binding to the Gla (gamma- carboxyglutamic acid) phospholipid binding domain of blood molecules with the structure determinations of PT Mg- and Tb-fragment1. These two molecules display deceptively similar properties to Ca-fragment1. However, our work of the past period suggests that the three most likely have very different three-dimensional structures. Diffraction quality single crystals of many of the above have been grown already and their X-ray diffraction patterns examined in a preliminary way.