NMR Investigation of Nucleotide Interactions with Cations and Proteins

核苷酸如三磷酸腺苷和GTP在许多生物活性中起着关键作用。核苷酸-阳离子 相互作用对于核苷酸的稳定、功能和调节是必不可少的。具体的和经常的 核苷酸与蛋白质的高度选择性相互作用是发挥其生物学功能所必需的 功能。这一建议旨在通过以下方法表征核苷酸与阳离子和蛋白质的相互作用 核磁共振波谱。建议的研究的具体目的是调查：(1) 核苷酸与阳离子的相互作用；(2)ATP/ADP与肌球蛋白相互作用的机制 肌动蛋白；(3)ATP/GTP与牛血清白蛋白(BSA)或泛素的结合。这些研究涉及核磁共振 化学位移、T1、T2弛豫和扩散测量。核磁共振数据将提供有价值的 资料：a)评估核苷酸的稳定性；b)关于GTP/ATP的水解机制；c) 评估核苷酸-肌球蛋白/肌动蛋白相互作用在ATP驱动的环结合解离中的作用 以及d)验证BSA和泛素是否是ATPase或 GTP酶诱导ATP或GTP水解酶。这些核磁共振研究具有重要的生物学意义和应用价值。 生物医学科学。PI为其MBRS-SCORE计划试点进行的成功研究- 使用萨凡纳州立大学核磁共振设施的项目(2001-2005年)构成了拟议的 研究。如果得到资助，这个成绩研究项目将使PI不仅能够继续他已经 建立了涉及核磁共振研究的研究，但也培训了MARC U*STAR和NSFHBCU的学生- UP程序。这项研究的结果将用于在国内和国际上展示 会议，在同行评议的期刊上发表，并寻求外部资金，以继续 超出MBRS-SCORE计划期的研究。

Nucleotides such as ATP and GTP play a pivotal role in many biological activities. Nucleotide-cation interactions are essential for the stabilization, function and regulation of nucleotides. The specific and often highly selective interactions of nucleotides with the proteins are required to perform their biological functions. This proposal aims at characterizing the interactions of nucleotides with cations and proteins by NMR spectroscopy. The specific aims of the proposed research are to investigate: (1) the interactions of nucleotides with cations; (2) the mechanisms involved in the interactions of ATP/ADP with myosin and actin; and (3) the binding of ATP/GTP to bovine serum albumin (BSA) or ubiquitin. The studies involve NMR chemical shifts, T1, T2 relaxation and diffusion measurements. The NMR data will provide valuable information: a) to assess the stability of nucleotides; b) on the mechanisms of GTP/ATP hydrolysis; c) to evaluate the interplay of nucleotide-myosin/actin interactions in the ATP-driven cyclic associationdissociation of actin-myosin; and d) to verify whether BSA and ubiquitin are the candidates as ATPase or GTPase to induce ATP or GTP hydrolysis. These NMR studies have significance in biological and biomedical sciences. The successful studies conducted by the PI for his MBRS-SCORE Program Pilot- Project (2001-2005) using NMR facility at Savannah State Univeristy formed the basis for the proposed research. If funded, this SCORE research project would enable the PI not only to continue his already establshied research involving NMR studies but also train students from the MARC U*STAR and NSFHBCU- UP Programs. The results from this study will be used for presenting at national and international conferences, for publication in peer-reviewed journals and for seeking extramural funding to continue the research beyond the MBRS-SCORE Program period.