ROLE OF Y-CARBOXYGLUTAMIC ACID IN BLOOD COAGULATION

Y-羧基谷氨酸在凝血中的作用

• 批准号: 3336052
• 负责人: RICHARD G HISKEY
• 金额: $ 24.16万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-12-01 至 1986-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3336052
• 关键词: arginine; blood coagulation; blood coagulation tests; calcium; chemical binding; chemical structure function; circular dichroism; coagulation factor IX; coagulation factor VII; coagulation factor X; congenital blood protein disorder; cystine; fluorimetry; gel filtration chromatography; hemophilia B; human subject; malonates; molecular pathology; nuclear magnetic resonance spectroscopy; nutrition related tag; phospholipids; protein C; prothrombin; radiotracer; vitamin K

The calcium ion binding and phospholipid binding behavior of prothrombin necessary for optimal rates of thrombin production in the blood coagulation process depend on the presence of gamma-carboxyglutamic acid (Gla) residues. This vitamin K-dependent amino acid is located at ten positions within the first 33 residues of prothromibin and at similar positions in the other vitamin K-dependent coagulation proteins factors VII, IX, X and Protein C. A related gamma-carboxyglutamix-containing region also occurs in bone proteins. We suggest that the 1-40 region of prothrombin, containing the 18-23 cystine loop, is critical to the functional behavior of prothrombin as well as related structures of the other vitamin K-dependent coagulation factors and perhaps other proteins, such as the bone protein, containing Gla and a similar cystine loop. We intend to prepare segments of the 1-40 region by chemical synthesis; we will also utilize the peptides representing the 1-39 and 12-44 sequences obtained from prothrombin. We intend to establish; (a) the chemical structure of the smallest unit of the amino terminal region of prothrombin that will exhibit the metal ion-induced characteristics of the protein; (b) the role of the cystine loop; (c) the nature of the ligand system in the Gla region that is required for metal ion binding; and, (d) the minimum number and location of Gla residues in the 1-33 sequence necessary for prothrombin-phospholipid interaction. Experimental techniques that will be employed to study the chemical and physical properties of these synthetic and natural peptides include nuclear magnetic resonance, laser Raman and Emu3+ luminescence spectroscopy; metal ion binding; interaction with antibodies specific to the calcium ion-induced conformation; and peptide; solvent and peptide:lipid partition experiments. A method for chemical modification of Gla residues is also proposed.

凝结凝结蛋白的钙离子结合和磷脂结合行为 血液凝结中凝血酶产生的最佳速率所必需的 过程取决于γ-羧基谷氨酸（GLA）的存在 残留物。 这种维生素K依赖性氨基酸位于十个位置 在丙光蛋白的前33个残留物中，在类似位置 其他维生素K依赖性凝血蛋白因子VII，IX，X和 蛋白质C.一个相关的含γ-羧基谷氨酸的区域也发生了 在骨蛋白中。 我们建议凝血酶原的1-40区域， 包含18-23的胱氨酸环，对功能行为至关重要 凝血酶原以及其他维生素的相关结构 K依赖性的凝血因子以及其他蛋白质，例如 骨蛋白，包含GLA和类似的胱氨酸环。 我们打算 通过化学合成制备1-40区域的段；我们也会 利用代表获得的1-39和12-44序列的肽 来自凝血酶原。 我们打算建立； （a）化学结构 凝血酶原氨基末端区域的最小单位 表现出金属离子诱导的蛋白质特征； （b）角色 胱氨酸循环； （c）GLA区域配体系统的性质 这是金属离子结合所必需的； （d）最小数字和 GLA残基在1-33序列中所需的位置 凝血酶原磷脂相互作用。 实验技术将是 用于研究这些合成的化学和物理特性 天然肽包括核磁共振，激光拉曼和 EMU3+发光光谱；金属离子结合；与 对钙离子诱导的构象特异的抗体；和肽； 溶剂和肽：脂质分区实验。 化学方法 还提出了GLA残基的修饰。