Study of Allosteric Proteins by NMR

描述（由申请人提供）：变构的原子尺度描绘 机制对理解生物分子功能做出了很大贡献。 Zuiderweg 博士已获得初步数据，使我们能够通过 溶液中的核磁共振波谱 (NMR)、变构 Hsp70 蛋白的机制。 NMR 能够整合以下研究： 结构、动力学和相互作用，因此可能有助于 对变构的基本理解，迄今为止几乎已经被 完全源自嵌入蛋白质结构的比较 晶体。 Zuiderweg 博士选择 Hsp70 伴侣蛋白系统作为 他的研究目标，因为其变构机制目前尚不清楚。 Hsp70 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。了解这些功能的功能 因此，分子与蛋白质疗法的开发具有相关性 折叠疾病。采用新开发的 NMR 方法，如 TROSY、光谱 通过氘化和特定标记进行简化以及测量 残余偶极耦合，目前可以研究大蛋白质 原子分辨率的解决方案。因此，通过 NMR 研究变构蛋白 已经触手可及；他的目标是 55 kDa。这是第一个结构研究 具有变构功能的 Hsp70 蛋白将有助于描述 控制之间变构耦合的构象/动力学变化 核苷酸和底物结合域。通过 NMR，可以研究这些 溶液的变化，并监测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 和钾。为了做到这一点，Zuiderweg 博士将首先集中精力 44 kDa 核苷酸结合域特性的 NMR 描述。在 下一阶段，Zuiderweg 博士将继续进行 55 kDa 构建，并研究 其分子参数作为核苷酸和底物结合的函数 合并。为了促进这项任务，Zuiderweg 博士的目标是 嗜热细菌 Dnak 伴侣结构的研究 嗜热栖热菌，可以在高温下进行研究，因此 产生出色的核磁共振谱。

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.