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.

我们实验室开发了大肠杆菌表达系统 诱导产生重组新型突变血红蛋白 位点特异性诱变。 该系统的目标之一是设计 突变体将允许我们在核磁共振谱中分配共振峰 人类血红蛋白。 因此，我们设计并生产了表面a-His 杜松子酒突变血红蛋白使我们能够分配一些芳香 共振。 一些突变血红蛋白显示出含氧量减少 合作性，表明显着的结构扰动 血红蛋白四聚体的亚基间区域。 这很有趣， 由于氨基酸取代位于血红蛋白的表面， $sim$30 $AA$ 距离子单元间区域太远，任何人都无法到达 怀疑血红蛋白界面有如此显着的影响 四聚体。 我们想研究单个氨基酸如何 表面的取代会扰乱血红蛋白的内部位点。 我们建议使用分子动力学来模拟结构 变化。 我们计划使用随机边界分子动力学 方法（SBMD）进行模拟，以减少处理时间。 我们可能 必须解决在计算中包括整个蛋白质， 然而，由于很难设计一个包含以下内容的边界： 表面的突变位点和原子的突变位点 亚基间表面。