BLOOD COAGULATION PROTEIN-METAL ION-LIPID INTERACTIONS

磷脂囊泡的相互作用模式的阐明， 膜模型，与金属离子，和各种凝血蛋白，和 这些相互作用导致酶原激活的方式， 参与血凝块的形成一直是一个长期的目标 这个实验室。 为了继续开发这一领域，我们将确定 一价和二价阳离子结合位点相互作用的方式 相互作用并与活化蛋白C中的活性位点结合;一种酶 这是一种有效的抗凝血剂。 一个强大的物理和 化学技术，如核磁共振和电子顺磁共振，将被用来解决这些问题 问题，这些问题将通过准备和 高度特异性化学试剂，单克隆抗体， 为了标记和鉴定蛋白质的参与其 许多功能。 将利用蛋白C的一种形式， “Gla-无结构域”-蛋白C，其缺乏许多阳离子结合位点， 天然蛋白C，以评估这种结构-功能关系 蛋白 将对凝血因子进行类似研究 IX和X。 通过多蛋白复合物激活因子X，还含有 钙和磷脂是血液凝固的中心环节。 在 为了确定这一过程的机制，我们将研究 条件下，物理和动力学，在此条件下，最佳激活 发生。 这些方法将再次通过准备和 单克隆抗体的表征，可能抑制或加速 在这个过程中的某些步骤，其中的识别将大大有助于 我们对这个系统的理解。 目前，人们对控制因素有很大的兴趣。 血栓形成和止血及其在生化水平上的阐明 肯定会建议操纵这个系统的手段， 人的优势。

The elucidation of the mode of interaction of phospholipid vesicles, as membrane models, with metal ions, and various coagulation proteins, and the manner in which these interactions lead to activation of zymogens which participate in formation of the blood clot has been a long-term objective of this laboratory. To continue to develop this area, we will determine the manner in which monovalent and divalent cation binding sites interact with each other and with the active site in activated protein C; an enzyme which is a potent anticoagulant. A powerful combination of physical and chemical techniques, such as NMR and EPR, will be employed to address these questions, and these will be extended by the preparation and characterization of highly specific chemical agents, monoclonal antibodies, to label and identify the portion of the protein which is involved in its many functions. Advantage will be taken of a form of protein C, "Gla-domainless"-protein C, which lacks many of the cation binding sites of native protein C, to assess structure-function relationships of this protein. Similar studies will be conducted with blood coagulation Factors IX and X. The activation of Factor X, by a multi-protein complex, also containing calcium and phospholipid, is a central event in the clotting of blood. In order to define the mechanism of this process, we will investigate conditions, both physically and kinetically, under which optimal activation occurs. These approaches will again be extended by the preparation and characterization of monoclonal antibodies, which may inhibit or accelerate certain steps in the process, the identification of which will greatly aid our understanding of this system. There is currently a great deal of interest in the factors which control thrombosis and hemostasis and their elucidation at the biochemical level will certainly suggest means of manipulation of this system to the advantage of man.