核磁共振波谱研究蛋白质三维结构及功能

Protein structure and function studied by NMR

文中总结了中国科学技术大学生命科学学院核磁共振波谱实验室十多年来的工作.我们的研究主要集中于研究人和其他真核生物基因表达调控相关蛋白质以及细胞连接处相关蛋白质.在这两个体系中许多蛋白质与人类健康及疾病相关,有的可能是潜在的药物作用靶标.我们主要关注用核磁共振波谱方法研究蛋白质-蛋白质相互作用的结构基础.核磁共振适合研究在接近生理条件下的分子相互作用,特别是适合研究低亲和力的瞬态的复合物.它可以提供蛋白质相互作用界面,复合物结构,以及蛋白质相互识别过程动力学的信息.文中给出了一些例子.我们也研究蛋白质内部动力学,包括皮秒-纳秒时间尺度,与毫秒-微秒时间尺度的动力学.与圆二色谱及荧光光谱结合,核磁共振可以详细表征蛋白质的折叠与去折叠.文中给出的核磁共振应用的最后一个例子是用计算机虚拟筛选,核磁筛选,我们发现了一个人的双功能的磷酸酶的一种新类型的抑制剂,并研究了该抑制剂对细胞功能的影响.这一策略有可能用于早期药物的发现.

More than 10 years of research in the Laboratory of Nuclear Magnetic Resonance （NMR） at the School of Life Science, University of Science and Technology of China is reviewed. Our researches have focused on two systems： proteins related to the regulation of gene expression in humans and other eukaryotes, and proteins existing in the cell junction in humans. The majority of proteins selected from these two systems are related to human health and diseases, and some are potential drug targets. We were interested in using NMR to study structural basis of protein-protein interactions. NMR was highly suited for investigating molecular interactions under approximately physiological conditions and was particularly suited for the study of low-affinity, transient complexes. It can provide information of protein interaction surface, complex structure, and dynamic properties during protein recognition. Several examples were given in this paper. NMR was also used to study dynamic properties of protein both in pico second to nano second and in micro-second to mili second time scales. We have studied protein folding and unfolding by NMR together with fluorescence and circular dichroism experiments. Proteins in unfolded states were characterized in detail by NMR. The last example of NMR application is the identification of a novel inhibitor of a human dual-specific phosphatase and the cellular effects of this compound were also studied. Our results demonstrate that our screening strategy, which combines both virtual and NMR-based methods, is feasible and might be employed in the early stage of drug discovery.