Studies of a thermophilic chaperonin system

嗜热伴侣蛋白系统的研究

• 批准号: 7383894
• 负责人: EDA KOCULI
• 金额: $ 2.56万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-10-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7383894
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Alzheimer' s Disease; Amyloidosis; Binding; Binding Proteins; Bovine Spongiform Encephalopathy; Cells; Cellular Membrane; Class; Complex; Cystic Fibrosis; DNA; Dependence; Disease; Escherichia coli; Fellowship; Goals; Heating; Hereditary Disease; In Vitro; Lead; Mediating; Molecular Chaperones; Nature; Numbers; Organism; Pathway interactions; Pharmaceutical Preparations; Play; Prion Diseases; Process; Protein Analysis; Protein Binding; Proteins; Proteomics; Recovery; Reporting; Role; Set protein; Stress; System; Temperature; Thermus thermophilus; chaperonin; expectation; extreme thermophile; insight; novel therapeutics; protein folding; protein function; protein misfolding; thermophilic organism

DESCRIPTION (provided by applicant): Protein folding is critical to the survival of all cells; it is the final step in the pathway from DNA to active protein. Proteins use various mechanisms to achieve their final folded state, including spontaneous folding and folding dependent on cellular machines known collectively as chaperones. The class of chaperones called chaperonins assists protein folding in an ATP-dependent manner and is responsible for folding many proteins, both newly synthesized and newly translocated across cellular membranes. They play a role in refolding heat or stress damaged proteins in the recovery from thermal or other insults. In this project, chaperonin-mediated protein folding by the GroEL-GroES system of the extreme thermophile, Thermus thermophilus, will be studied with the long-term goal of understanding how cellular systems have evolved to deal with environmental extremes and, more specifically, what the differences are between folding mechanisms in mesophilic organisms and those in thermophiles. The specific aims of this project are to: 1) identify potential substrate proteins for the thermophilic chaperonin, in addition to the 24 reported recently, by proteomic analysis of the protein occupants of the chaperonin complex recovered from T. thermophilus cells and evaluate both sets of proteins for their dependence on the chaperonin system for refolding in vitro, examining aggregation and chaperonin binding when diluted from denaturant, release by the addition of ATP and co-chaperonin, and recovery of native enzymatic activity or protein function; 2) investigate the ATPase and protein folding cycles of the T. thermophilus chaperonin system and determine the efficiency of refolding in terms of cycles of ATP hydrolysis required to fold a given protein, comparing this to the efficiency of the well-studied E. coli chaperonin system, both with homologous substrate proteins and with identical mesophilic or thermophilic proteins at the same temperature; and 3) initiate NMR studies of a thermophilic substrate protein bound to thermophilic GroEL, with the expectation that the ability to collect spectra at elevated temperature may provide new insights into the nature of the bound protein and its interaction with the chaperonin. Not only is the description of how proteins fold central to our understanding of how cells survive and grow, it is crucial to uncovering the causes behind diseases that result from protein misfolding. These include the amyloidoses, such as Alzheimer disease, prion diseases, such as "mad cow" disease, and certain genetic diseases, such as the most common form of cystic fibrosis. Understanding the processes of protein folding may lead to the uncovering of new drugs and new therapeutic approaches to these severe disorders. Page 2 Number pages consecutively at the bottom throughout Form Page 2

描述（由申请人提供）：蛋白质折叠对所有细胞的存活至关重要;它是从DNA到活性蛋白质途径的最后一步。蛋白质使用各种机制来实现其最终折叠状态，包括自发折叠和依赖于统称为分子伴侣的细胞机器的折叠。称为伴侣蛋白的一类伴侣蛋白以ATP依赖性方式协助蛋白质折叠，并负责折叠许多新合成的和新跨细胞膜转运的蛋白质。它们在从热或其他损伤中恢复的热或应力损伤蛋白质的重折叠中起作用。在这个项目中，伴侣蛋白介导的极端嗜热菌，嗜热栖热菌的GroEL-GroES系统的蛋白质折叠，将与了解细胞系统如何演变来应对环境极端的长期目标进行研究，更具体地说，在嗜温生物和嗜热生物的折叠机制之间有什么区别。本项目的具体目标是：1）通过对从T. 2）研究了嗜热细胞的ATP酶和蛋白质折叠循环; 3）研究了嗜热细胞的ATP酶和蛋白质折叠循环。嗜热菌伴侣蛋白系统，并确定折叠给定蛋白质所需的ATP水解循环的重折叠效率，将其与充分研究的E. coli伴侣蛋白系统，在相同温度下具有同源底物蛋白和相同的嗜中温或嗜热蛋白;和3）启动与嗜热GroEL结合的嗜热底物蛋白的NMR研究，期望在升高的温度下收集光谱的能力可以提供对结合蛋白的性质及其与伴侣蛋白的相互作用的新见解。蛋白质如何折叠的描述不仅是我们理解细胞如何生存和生长的核心，而且对于揭示蛋白质错误折叠导致的疾病背后的原因至关重要。这些包括淀粉样变性，如阿尔茨海默病，朊病毒疾病，如“疯牛病”，和某些遗传疾病，如最常见的囊性纤维化。了解蛋白质折叠的过程可能会导致这些严重疾病的新药和新的治疗方法的发现。第2页在表格第2页的底部连续编号