Dissecting the structure and function of the Pdu microcompartment in Salmonella

解析沙门氏菌 Pdu 微区室的结构和功能

• 批准号: 7895697
• 负责人: THOMAS Aquinas BOBIK
• 金额: $ 54.57万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895697
• 关键词: Affect; Architecture; Bacteria; Biochemical; Biological Assay; Cell physiology; Comparative Study; Complex; Computing Methodologies; Crystallography; Cytosol; Encapsulated; Environment; Enzymes; Face; Genes; Genetic; Goals; Growth; Higher Order Chromatin Structure; Human; Hybrids; Immunoelectron Microscopy; In Vitro; Individual; Investigation; Knowledge; Label; Lead; Link; Maps; Mediating; Metabolic; Metabolic Pathway; Metabolism; Methods; Microbe; Molecular; Mutagenesis; Mutation; Organelles; Pathogenesis; Play; Process; Production; Propylene Glycols; Proteins; Publishing; Reaction; Relative (related person); Research; Role; Salmonella; Salmonella enterica; Salmonella typhimurium LT2; Side; Solutions; Spatial Distribution; Structure; Subcellular structure; Surface; System; Testing; Time; Toxic effect; Transport Process; Virus; Work; X-Ray Crystallography; cofactor; enzyme substrate; in vivo; insight; mutant; novel; pathogen; polypeptide; prevent; protein protein interaction; protein structure; public health relevance; structural biology; three dimensional structure; tool; yeast two hybrid system

DESCRIPTION (provided by applicant): Bacterial microcompartments are large subcellular structures composed of metabolic enzymes encapsulated within a protein shell built from multiple subunits. They are widespread among bacteria, functionally diverse, play vital metabolic roles, are linked to pathogenesis, and appear to incorporate unique mechanistic and structural principles. Their function is to sequester and regulate the production of toxic or volatile intermediates found in certain metabolic pathways. However, little is known about how this is occurs at the mechanistic level. The long-term goal of the proposed research is to elucidate the molecular principles and to build up a 3-dimensional structure of the microcompartments involved in 1,2-propanediol degradation by Salmonella. The Salmonella system is unmatched with regard to the knowledge and tools available for mechanistic studies of microcompartments. The proposed studies combine genetic, biophysical, and structural methods to elucidate the cellular function of the Salmonella Pdu microcompartment at a mechanistic level. Three specific aims are proposed: 1. determine the structures of the microcompartment shell proteins and enzymes; 2. elucidate biochemical interactions and higher-order architecture in the Pdu microcompartment; and 3. conduct mutational analysis of microcompartment function. Structures will be investigated and analyzed by x-ray crystallography, biophysical, and computational methods. Interactions studies will include two-hybrid analyses, labeling studies, and crystallography. Functional and mechanistic insights will be derived from structure-guided mutagenesis in conjunction with growth studies, enzyme and transport assays, and reverse two-hybrid analyses. Completion of the proposed investigations will elucidate the mechanistic and structural principles of the Salmonella pdu microcompartment. This will provide general insights into bacterial microcompartments. Furthermore, since bacterial microcompartments play critical metabolic roles in many microbes, including several human pathogens, the proposed studies may ultimately lead to new opportunities for interfering with pathogenic processes. PUBLIC HEALTH RELEVANCE: Bacterial microcompartments play critical metabolic roles in many microbes, including several human pathogens, but they are very poorly understood at the present time. This proposal seeks to understand how the Pdu microcompartment functions in the human pathogen, Salmonella enterica serovar Typhimurium. The studies will lead to a deeper fundamental and mechanistic understanding of these important bacterial organelles, and ultimately to new opportunities for interfering with pathogenic processes.

描述（由申请人提供）：细菌微校区是大型亚细胞结构，该结构由封装在由多个亚基建造的蛋白质壳中的代谢酶组成。它们在细菌中普遍存在，功能多样，扮演重要的代谢角色，与发病机理有关，并且似乎结合了独特的机械和结构原理。它们的功能是隔离和调节在某些代谢途径中发现的有毒或挥发性中间体的产生。但是，关于这种情况如何在机械水平上发生的知之甚少。拟议研究的长期目标是阐明分子原理，并建立三维结构的三维结构，使沙门氏菌涉及1,2-丙二醇降解。关于用于微校区的机械研究的知识和工具，沙门氏菌系统是无与伦比的。拟议的研究结合了遗传，生物物理和结构方法，以在机械水平上阐明沙门氏菌PDU微室的细胞功能。提出了三个特定的目的：1。确定微木壳蛋白和酶的结构； 2。阐明PDU微校区中的生化相互作用和高阶结构；和3。进行微校区函数的突变分析。结构将通过X射线晶体学，生物物理和计算方法研究和分析。相互作用的研究将包括两个杂交分析，标记研究和晶体学。功能和机械洞察力将来自结构引导的诱变，并结合生长研究，酶和运输分析，并逆转两种杂交分析。拟议的调查的完成将阐明沙门氏菌PDU微校区的机械和结构原理。这将提供对细菌微型营的一般见解。此外，由于细菌微校区在包括几种人类病原体在内的许多微生物中起关键的代谢作用，因此拟议的研究最终可能会带来新的机会来干扰致病过程。公共卫生相关性：细菌微型院子在许多微生物（包括几种人类病原体）中扮演着重要的代谢角色，但目前对它们的了解很少。该提案旨在了解PDU微室在人类病原体Salmonella enterica serovar typhimurium中的功能。这些研究将导致对这些重要细菌细胞器的更深入和机械理解，并最终带来干扰致病过程的新机会。