Study of Allosteric Proteins by NMR

描述（由申请人提供）：Hsp70（热休克70 kDa）伴侣蛋白在正常和应激细胞条件下，从古生菌到智人的生物中都是蛋白质折叠、再折叠和运输的核心。Hsp70's （re）通过底物结合域的结合和释放循环折叠蛋白质，涉及ATP和ADP在核苷酸结合域的结合引起的构象变化。这种远程调节机制被称为变构。最近，热休克蛋白70与乳腺癌等疾病有关。用小化合物调节Hsp70的变构机制可能形成治疗这些疾病的途径。我们建议通过确定60 kDa Hsp70蛋白结构体的溶液构象来详细研究其机制，这些结构体包含核苷酸和底物结合结构域以及它们与不同配体状态的共伴侣的复合物。我们将研究第一代Hsp70调节化合物作用的分子基础。所获得的综合见解将有助于设计用于治疗细胞管理疾病的改进化合物。这项研究将使用超高场核磁共振波谱进行。

DESCRIPTION (provided by applicant): Hsp70 (heat shock 70 kDa) chaperone proteins are central to protein folding, refolding, and trafficking in organisms ranging from Archae to Homo Sapiens, both at normal and at stressed cellular conditions. Hsp70's (re) fold proteins via binding and release cycles at the substrate-binding domain involving conformational changes caused by ATP and ADP binding at the nucleotide-binding domain. This remote regulation mechanism is called allostery. Recently, Hsp70's have been linked to diseases such as breast cancer. Modulation of the allosteric mechanism of the Hsp70's with small compounds may form an avenue to treat these diseases. We propose to investigate the mechanism in detail by determining the solution conformations of 60 kDa Hsp70 protein constructs, containing both nucleotide- and substrate-binding domains and their complexes with co-chaperones in different liganded states. We will investigate the molecular basis for the action of the first generation of Hsp70 modulating compounds. The combined insights gained will aid in the design of improved compounds for the treatment of cell-management diseases. The research will be carried out using ultra-high field nuclear magnetic resonance spectroscopy.