NMR INVESTIGATION OF HUMAN CKSHS1

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。项目名称：人类CksHs1核磁共振研究项目负责人：James J.A. Huntley博士工作地点：国家基因组资源中心，斯克里普斯研究所，新墨西哥州立大学这项研究的广泛而长期的目标是确定蛋白质结构动力学在构成细胞生长和分裂过程中的作用。这项研究的具体目的是探索和表征一种被认为具有多种作用的蛋白质的动力学特性，包括一种有助于加强细胞周期的正确时间和进程的蛋白质。这种名为CksHs1的蛋白已被证明与许多蛋白质和无机分子结合，包括磷酸盐和细胞周期蛋白依赖性激酶2 (CDK2)，一种参与细胞周期调节的关键酶。CksHs1在不同状态下的比较表明，蛋白质的构象灵活性是其功能的一个重要方面。然而，这些研究并没有完全描述这种灵活性，或者它与CksHs1功能的确切关系。没有这些信息，就不可能全面了解CksHs1在细胞生长和分裂过程中的功能。为了更好地理解动力学在CksHs1功能中的作用，我们将利用核磁共振弛豫技术研究CksHs1的结构。我们的中心假设是CksHs1表现出新的动力学特征，使蛋白质能够根据结合伙伴的结构折叠成略有不同的结构。我们的目标是表征游离CksHs1的动力学，CksHs1在与磷酸盐结合时发生的动力学变化，绘制CksHs1和CDK2相互作用的图谱，并表征CksHs1-CDK2结合过程的动力学。这项研究的基本原理是，我们将更好地了解细胞周期控制的组成部分和途径，以及最终如何利用蛋白质结构动力学的特征来开发新的治疗药物或策略来治疗癌症或其他细胞生长不受控制的疾病。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. TITLE OF PROJECT: NMR Investigation of Human CksHs1 PILOT-PROJECT INVESTIGATOR: James J.A. Huntley, Ph.D. PERFORMANCE SITES: National Center for Genome Resources, The Scripps Research Institute, New Mexico State University The broad, long-term objective of this investigation is to determine the role of protein structural dynamics in the processes that constitute cell growth and division. The investigation¿s specific aim is to explore and characterize the dynamics of a protein believed to have multiple roles, including one that serves to reinforce the correct timing and progression of the cell cycle. The protein, CksHs1, has been shown to bind to a number of proteins and inorganic molecules, including phosphate and Cyclin Dependent Kinase 2 (CDK2), a key enzyme involved in cell cycle regulation. Comparison of CksHs1 in various states has suggested that conformational flexibility of the protein is an important facet of its function. However, these investigations have not fully characterized this flexibility, or exactly how it relates to CksHs1 function. Without this information, a complete picture of CksHs1 function in the process of cell growth and division will not be possible. In order to better understand the role of dynamics in CksHs1 function, we will study the structure of CksHs1 by use of Nuclear Magnetic Resonance (NMR) relaxation techniques. Our central hypothesis is that CksHs1 displays novel features of dynamics that give the protein the ability to fold into slightly different structures depending upon the structure of the binding partner(s). We aim to characterize the dynamics of free CksHs1, the alteration to CksHs1 dynamics that occurs upon binding of phosphate, map CksHs1 and CDK2 interactions, and characterize the dynamics of the CksHs1-CDK2 binding process. The rationale for this research is that we will better understand the components and pathways of cell cycle control, and ultimately how features of protein structure dynamics may be exploited in the development of novel therapeutic agents or strategies to treat cancer or other diseases of uncontrolled cell growth.