DYNAMICS OF GLUTATHIONE TRANSFERASE A1-1

谷胱甘肽转移酶 A1-1 的动力学

• 批准号: 7723378
• 负责人: GORDON RULE
• 金额: $ 0.05万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723378
• 关键词: Binding; Complement; Complex; Computer Retrieval of Information on Scientific Projects Database; Condition; Docking; Drug Metabolic Detoxication; Enzymes; Funding; Future; Glutathione; Glutathione S-Transferase; Grant; Helix (Snails); Homo; Housing; Human; Institution; Laboratories; Ligands; Molecular Weight; Mutation; NMR Spectroscopy; Numbers; Outcome; Property; Protein Dynamics; Protein Isoforms; Proteins; Research; Research Personnel; Resources; Running; Services; Source; System; United States National Institutes of Health; alpha helix; dimer; ear helix; enzyme structure; glutathione transferase A1-1; molecular mechanics; mutant; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Glutathione transferases (GST) are detoxification enzymes that inactivate reactive hydrophobic compounds by conjugating the tripeptide glutathione to reactive centers on these compounds. We have been using NMR spectroscopy to characterize the dynamic properties of one human isoform, GST A1-1. We have shown that the carboxy-terminal alpha helix becomes docked when either substrate or product bind to the enzyme. Currently we are generating mutations in the protein that may be responsible for stabilizing the docked form of this helix and investigating the enzymatic and dynamic properties of these enzymes. To complement the NMR studies, we would like to investigate the effects of these mutations on the dynamics of the protein using molecular mechanics calculations. We feel that the molecular mechanics calculations will aid in the interpretation of our laboratory date and may provide a means to predict the outcome of future experiments. The protein is a homo-dimer of 50,000 kDa overall molecular weight. We would like to perform dynamics calculations on the wild-type enzyme and three mutants (Ile219Ala, Phe220Ala, Phe222Ala, for both the unliganded and the enzyme-product complex. The crystal structure of the enzyme-product complex will be used for starting coordinates. We intend to employ CHARMM using a solvated system with periodic boundary conditions. We will initially perform dynamics runs for ~5ns to gauge whether there are significant difference between the eight different forms of the protein (apo + liganded/wildtype + 3 mutants). We anticipate equilibration of the system in-house on our sgi origin 300 server. Please allocate the appropriate number of service units.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 谷胱甘肽转移酶 (GST) 是一种解毒酶，可通过将三肽谷胱甘肽缀合到这些化合物的反应中心来灭活反应性疏水化合物。我们一直在使用核磁共振波谱来表征一种人类亚型 GST A1-1 的动态特性。我们已经证明，当底物或产物与酶结合时，羧基末端α螺旋就会对接。目前，我们正在蛋白质中产生突变，这些突变可能负责稳定该螺旋的对接形式，并研究这些酶的酶学和动态特性。为了补充核磁共振研究，我们希望使用分子力学计算来研究这些突变对蛋白质动力学的影响。我们认为分子力学计算将有助于解释我们的实验室数据，并可能提供预测未来实验结果的方法。该蛋白质是总分子量为 50,000 kDa 的同源二聚体。我们想要对野生型酶和三个突变体（Ile219Ala、Phe220Ala、Phe222Ala，对于未配体和酶-产物复合物）进行动力学计算。酶-产物复合物的晶体结构将用于起始坐标。我们打算使用具有周期性边界条件的溶剂化系统的 CHARMM。我们将首先进行动力学运行 约 5ns 来衡量八种不同形式的蛋白质（apo + 配体/野生型 + 3 个突变体）之间是否存在显着差异。我们预计 sgi origin 300 服务器上的内部系统会达到平衡。请分配适当数量的服务单位。