Studies of a thermophilic chaperonin system

嗜热伴侣蛋白系统的研究

基本信息

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
项目类别：
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

项目摘要

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）：蛋白质折叠对于所有细胞的存活至关重要；这是从DNA到活性蛋白的途径的最后一步。蛋白质使用各种机制实现其最终折叠状态，包括自发折叠和折叠，取决于统称为伴侣的细胞机。称为伴侣蛋白的伴侣类别以ATP依赖性的方式有助于蛋白质折叠，并负责折叠许多蛋白质，这些蛋白质是新合成的，既新合成又跨细胞膜。它们在从热或其他侮辱中恢复中恢复热量的蛋白质中发挥作用。在这个项目中，将研究伴侣蛋白介导的蛋白质折叠，该蛋白质由极端热嗜热嗜热嗜热的gro折叠，其长期目标是理解细胞系统如何演变为如何发展为环境极端的发展，更具体地，具体地，差异是在细胞生物体中的折叠机制之间的差异和热量剂中的差异。该项目的具体目的是：1）通过蛋白质组学分析，除了最近报道的24种报道外，除了蛋白质组占用者外，还确定了嗜热伴侣蛋白的潜在底物蛋白，除了从T. thepophilus细胞中恢复的蛋白质组占用物的蛋白质组占用物，并评估蛋白质对蛋白质的依赖性，并评估蛋白质对蛋白质的依赖，并在蛋白质中弥补蛋白质，并延伸了依然的蛋白质，并延伸了依然的蛋白质。通过添加ATP和共伴侣蛋白，并恢复天然酶活性或蛋白质功能； 2）研究T. hythphilus伴侣蛋白系统的ATPase和蛋白质折叠循环，并确定折叠给定蛋白需要的ATP水解循环的重折叠效率，将其与良好的大肠杆菌伴侣蛋白系统的效率进行比较，并将其与同源性蛋白质和识别蛋白质相同或热蛋白质蛋白质或热蛋白质相同； 3）启动与嗜热凹槽结合的嗜热底物蛋白的NMR研究，并期望在升高温度下收集光谱的能力可以为结合蛋白质的性质及其与伴侣蛋白的相互作用提供新的见解。不仅描述蛋白质如何折叠我们对细胞如何生存和生长的理解中心，而且至关重要的是要揭示蛋白质折叠折叠导致的疾病背后的原因。这些包括淀粉样蛋白，例如阿尔茨海默氏病，prion病，例如“疯牛”疾病和某些遗传疾病，例如囊性纤维化的最常见形式。了解蛋白质折叠的过程可能会导致新药物的发现以及这些严重疾病的新治疗方法。第2页数字页面在整个表格第2页的底部连续底部