Study of Allosteric Proteins by NMR

描述(申请人提供)：变构的原子尺度描述 机制对理解生物分子的功能有很大的贡献。 Zuiderweg博士已经获得了初步数据，将使我们能够通过 变构--溶液核磁共振波谱 Hsp70蛋白的作用机制。核磁共振能够整合对核磁共振的研究。 结构、动力学和相互作用，因此很可能有助于 对变构的基本理解，到目前为止，它几乎 独家来源于对嵌入的蛋白质结构的比较 水晶。Zuiderweg博士选择Hsp70伴侣蛋白系统作为 作为他研究的目标，因为其变构机制目前尚不清楚。 热休克蛋白70的S作为最丰富和最保守的系统发挥着核心作用 帮助蛋白质在体内折叠。对这些功能的理解 因此，分子对蛋白质疗法的发展具有相关性。 折叠疾病。用新开发的核磁共振方法，如TROSY，SPECTRUM 重氢和特定标记法的简化及其测量 残留的偶极偶联，目前有可能研究大的蛋白质 原子分辨率的溶液。因此，变构蛋白的核磁共振研究 已经触手可及；他的目标是55 kDa。这是第一次对An进行结构研究 变构功能的Hsp70蛋白将有助于描绘 控制变构偶联的构象/动力学变化 核苷酸结合区和底物结合区。通过核磁共振，有可能研究这些 溶液中的变化，并监控添加对这些参数的影响 不同的核苷酸、底物和辅因子，如磷酸盐、镁 还有钾。为了做到这一点，Zuiderweg博士将首先专注于 核磁共振描述44 kDa核苷酸结合域的性质。在 下一阶段，Zuiderweg博士将继续研究55 kDa的结构，并研究 其分子参数与核苷酸和底物结合的关系 加在一起。为了促进这项任务，Zuiderweg博士将致力于 嗜热菌Dnak分子伴侣结构的研究 嗜热性嗜热菌，可以在高温下研究，因此 产生了极好的核磁共振光谱。

DESCRIPTION (provided by applicant): The atomic-scale delineation of allosteric mechanisms has contributed much to the understanding of biomolecular function. Dr. Zuiderweg has obtained preliminary data that will allow us to study by nuclear magnetic resonance spectroscopy in solution (NMR), the allosteric mechanisms of Hsp70 proteins. NMR is capable of integrating the study of structure, dynamics and interactions and is therefore likely to contribute to the fundamental understanding of allosterics, which thus far has been almost exclusively derived from comparisons of structures of proteins embedded in crystals. Dr. Zuiderweg has chosen the Hsp70 chaperone protein system as a target for his studies because its allosteric mechanism is currently unknown. The Hsp70's play a central role as the most abundant and most conserved systems aiding protein folding in vivo. Understanding of the functioning of these molecules is thus of relevance for the development of therapies for protein folding diseases. With newly developed NMR methods such as TROSY, spectral simplification by deuteration and specific labeling and the measurement of residual dipolar couplings, it is currently possible to study large proteins in solution at atomic resolution. As such, the study of allosteric proteins by NMR has come within reach; his target is 55 kDa. This first structural study of an allosterically functional Hsp70 protein will help delineate the conformational/dynamical changes that govern the allosteric coupling between nucleotide and substrate-binding domains. By NMR, it is possible to study these changes in solution, and monitor the effects on these parameters of adding different nucleotides, substrates, and co-factors such as phosphate, magnesium and potassium. In order to do so, Dr. Zuiderweg will first concentrate on the NMR description of the properties of 44 kDa nucleotide binding domains. In the next stage, Dr. Zuiderweg will move onward to the 55 kDa construct, and study its molecular parameters as a function of nucleotide and substrate binding combined. In order to facilitate this task, Dr. Zuiderweg will aim for the study of such a construct of the Dnak chaperone of the thermophilic bacterium Thermus thermophilus, which can be studied at elevated temperatures and hence gives rise to excellent NMR spectra.