AUTOMATED INTERACTIVE MICROSCOPE HIGH PERFORMANCE IMAGING FOR BIOLOGIC RESEARCH

用于生物研究的自动交互式显微镜高性能成像

• 批准号: 6221061
• 负责人: F LANNI
• 金额: $ 0.13万
• 依托单位: MELLON PITTS CORPORATION (MPC CORP)
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6221061
• 关键词: bacteria; biological products; biomedical resource; computers; hematology

Our laboratory has developed an E. Coli expression system for production of recombinant novel mutant hemoglobins induced by site-specific mutagenesis. One objective of this system is to design mutants that will allow us to assign resonance peaks in NMR spectra of human hemoglobin. We have thus designed and produced surface a-His to Gin mutant hemoglobins that have enabled us to assign some aromatic resonances. Some mutant hemoglobins have shown decrease in oxygen cooperativity, suggesting significant structural perturbation at the intersubunit region of hemoglobin tetramer. This is interesting, since the amino acid substitution is on the surface of the hemoglobin, $sim$30 $AA$ away from the intersubunit region, too far for anyone to suspect such prominent influence at the interface of the hemoglobin tetramer. We would like to investigate how a single amino acid substitution at the surface perturbs the interior sites of hemoglobin. We propose to use molecular dynamics to simulate the structural changes. We plan to use the stochastic boundary molecular dynamics method(SBMD) for simulation to reduce the processing time. We may have to resolve to including the entire protein in the calculation, however, since it is difficult to design a boundary that will include both the mutation site at the surface and the atoms at the intersubunit surface.

我们的实验室已经开发了一个大肠杆菌表达系统 产生重组新型突变体血红蛋白。 位点特异性诱变。 该系统的一个目的是设计 将使我们能够在NMR光谱中分配共振峰的突变体 人血红蛋白。 因此，我们已经设计和生产了表面A-HIS 杜松子酒突变血红蛋白使我们能够分配一些芳香 共鸣。 一些突变的血红蛋白显示氧气减少 协作性，表明在 血红蛋白四聚体的亚基区。 这很有趣， 由于氨基酸取代在血红蛋白的表面 $ sim $ 30 $ aa $远离sububunit地区，任何人都太远了 怀疑血红蛋白界面的如此突出的影响 四聚体。 我们想研究一个单一氨基酸 在表面取代血红蛋白的内部部位。 我们建议使用分子动力学来模拟结构 更改。 我们计划使用随机边界分子动力学 方法（SBMD）进行仿真以减少处理时间。 我们可能 必须解决在计算中包含整个蛋白质的 但是，由于很难设计一个将包括的边界 表面的突变位点和原子 亚基间表面。