HOW SITE DIRECTED MUTANTS AFFECT HBS FIBER STRUCTURE

定点突变体如何影响 HBS 纤维结构

• 批准号: 6110863
• 负责人: ROBERT JOSEPHS
• 金额: --
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6110863
• 关键词: hemoglobin Ss; human tissue; intracellular; polymerization; protein structure function; site directed mutagenesis

In this project, Dr. Josephs proposes to extend his extensive and elegant earlier work, that has culminated in a highly detailed model for the structure of the sickle cell hemoglobin fiber. This model provides the foundation for understanding the interactions that drive fiber formation and, thus, the clinical manifestations of sickle-cell anemia. The fundamental unit of the sickle-cell fiber is a double-strand of hemoglobin molecules that has been observed in crystals of deoxy hemoglobin molecules S (HbS). The fiber is composed of seven of these double strands that have a helical twist in the fiber, but not in the crystal. The crystal structure analysis by Love and colleagues delineates the interactions within the double strand, however the interactions between double strands that stabilize the fiber are not present in the crystal lattice. Thus, EM analysis, in conjunction with model building, is required to determine the "inter double strand" contacts. Two competing models have been proposed- one by the group of Dr. Josephs and the other by Edelstein and colleagues. With the use of an efficient microbial systems for production of recombinant hemoglobin, these alternate models can now be tested by site- directed mutagenesis. The proposed project contains three specific aims.

在这个项目中，约瑟夫斯博士建议扩展他的广泛优雅 较早的工作，最终导致了一个高度详细的模型 镰状细胞血红蛋白纤维的结构。该模型提供了 理解驱动纤维形成的相互作用的基础 因此，镰状细胞贫血的临床表现。这 镰状细胞纤维的基本单位是血红蛋白的双链 在脱氧血红蛋白分子晶体中观察到的分子 S（HBS）。纤维由其中七个具有的双链组成 纤维中的螺旋扭转，但没有晶体。水晶 爱与同事的结构分析描述了互动 在双链中，但是双链之间的相互作用 稳定纤维不存在晶格中。因此，em 与模型构建结合进行分析以确定 “间双链”触点。已经提出了两个竞争模型 - 一个由约瑟夫斯博士组成，另一个由爱德尔斯坦和同事组成。 使用有效的微生物系统生产 重组血红蛋白，这些替代模型现在可以通过站点测试 定向诱变。拟议的项目包含三个具体目标。