ATPASE KINETICS AND MACROMOLECULAR ASSEMBLIES OF S54-DEPENDENT, AAA+ ATPASES

S54 依赖性 AAA ATP酶的ATP酶动力学和大分子组装

• 批准号: 8361268
• 负责人: B TRACY NIXON
• 金额: $ 0.59万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361268
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Behavior; Binding; Biophysics; Cell division; Cells; Cellular Structures; DNA; Defect; Disease; Funding; Gene Expression; Genetic Transcription; Grant; Human; Hydrogen; Kinetics; Measurement; Mechanics; Membrane; Modeling; Molecular Machines; Mutate; National Center for Research Resources; Nitrogen Fixation; Organelles; Principal Investigator; Production; Protein Binding; Proteins; Protomer; Publications; Research; Research Infrastructure; Resources; Source; Structure; United States National Institutes of Health; Work; cost; enhancer binding protein; macromolecular assembly; mutant; novel; pathogen; remediation; repaired

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Why do defects in molecular machines (AAA+ ATPases) cause disease? These protein machines convert ATP hydrolysis into mechanical work. Both human cells and disease causing pathogens use this work to physically manipulate proteins or DNA to dismantle and reassemble membranes or other organelles, to replicate DNA and traverse cell division, to repair damaged proteins, or to regulate gene expression. We do not know how these molecular machines convert ATP hydrolysis into mechanical work. Our research focuses on one subset of AAA+ ATPAses, the bacterial-enhancer-binding proteins (EBPs) which use their ATPase activities to regulate transcription of genes needed for harmful activites (diseases, crop damage) or helpful ones (nitrogen fixation, environmental remediation, hydrogen or other metabolite production). We are addressing the underlying mechanism of AAA+ ATPase function via structure function studies of mutant forms of ATPase. We have combined the functional studies with analysis of a new crystal structure for one of the mutant proteins bound to ATP. The analysis yielded a novel perspective of the conformational changes that occur upon the protomers moving from ATP-bound to ADP-bound. This model predicted unexpected residues would be important for function. We mutated one of the key residues, and used SAXS measurements to study its behavior in the presence of nucletotides. That work was just accepted for publication by Structure (Cell Press).

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 为什么分子机器(AAA+ATPase)中的缺陷会导致疾病？这些蛋白质机器将三磷酸腺苷的水解转化为机械功。人类细胞和引起疾病的病原体都利用这种工作来物理地操纵蛋白质或DNA，以拆解和重组膜或其他细胞器，复制DNA并穿越细胞分裂，修复受损的蛋白质，或调节基因表达。我们不知道这些分子机器是如何将三磷酸腺苷的水解转化为机械功的。我们的研究集中在AAA+ATPase的一个子集，细菌增强子结合蛋白(EBPs)，它利用它们的ATPase活性来调节有害活动(疾病、作物损害)或有用活动(固氮、环境修复、氢气或其他代谢物产生)所需基因的转录。 我们正在通过对ATPase突变形式的结构功能的研究来探讨AAA+ATPase功能的潜在机制。我们将功能研究与对其中一种与ATP结合的突变蛋白的新晶体结构的分析结合在一起。这一分析为原基从ATP结合到ADP结合时发生的构象变化提供了一个新的视角。该模型预测意外残基将对功能起重要作用。我们突变了一个关键残基，并使用SAXS测量来研究它在核存在下的行为。这部作品刚刚被Structure(Cell Press)接受出版。