Functional Dynamics of the Myosin Molecular Motor

描述(申请人提供)：申请者尤里·E·内斯梅洛夫博士是一名物理学家，他的研究重点是电磁场与物质的相互作用。内斯梅洛夫博士建议学习和利用分子生物物理和生化技术，以完善用于研究肌球蛋白功能动力学的多学科方法。职业发展/培训计划：该计划的主要内容将是从指导培训中获得先进的分子生物物理和生化技术。大部分培训将来自主要导师大卫·D·托马斯博士，他是肌肉生物物理学领域的国际知名科学家，也是NIH资助的一个肌肉研究培训项目的负责人。细胞和分子生物学方面的额外培训将由共同导师玛格丽特·蒂图斯博士提供，她因在肌球蛋白方面的工作而闻名。部分研究(W波段电子顺磁共振(EPR))将与合作者Peter Fajer博士一起进行，他是著名的生物EPR和肌肉生物物理学专家。为进一步发展职业和研究而组织的活动将包括出席和参加国家科学会议，定期参加期刊俱乐部和研讨会，以及加入明尼苏达大学肌肉营养不良中心。研究计划：这项建议的总体目标是让内斯梅洛夫博士学习分子生物物理和生化技术，并将它们与定点自旋标记和多频率EPR谱一起应用，以探索肌球蛋白运动功能的内部动力学。建议的工作是基于申请人在提高EPR测量灵敏度方面的最新进展，特别是在溶液中肌球蛋白的实验中，以及在多频EPR数据分析中，以准确表征蛋白结合自旋标记的动力学。这种方法将被用来获得关于蛋白质结构元件动态重排的详细信息。这些方法将被进一步发展，并用于确定肌球蛋白ATPase循环中肌球蛋白中继发螺旋和SH1螺旋的结构重排动力学。

DESCRIPTION (provided by applicant): The applicant, Dr. Yuri E. Nesmelov, is a physicist whose research has focused on the interaction of electromagnetic fields with matter. Dr. Nesmelov proposes to learn and utilize molecular biophysical and biochemical techniques to round out a multidisciplinary approach to be used in studying myosin functional dynamics. Career Development/Training Plan: The primary element of the plan will be the acquisition of advanced molecular biophysical and biochemical techniques from mentored training. The majority of the training will come from the primary mentor, Dr. David D. Thomas, who is an internationally known scientist in the field of muscle biophysics, and is the director of an NIH-funded training program in muscle research. Additional training in cell and molecular biology will be provided by the co-mentor, Dr. Margaret Titus, who is well known for her work on myosin. Part of the research (W band electron paramagnetic resonance (EPR)) will be performed with a collaborator, Dr. Peter Fajer, a renowned specialist in biological EPR and muscle biophysics. Structured activities for further career and research development will include attendance and participation at national scientific meetings, regular involvement in journal clubs and seminars, and affiliation with the Muscular Dystrophy Center at the University of Minnesota. Research Plan: The overall aim of this proposal is for Dr. Nesmelov to learn molecular biophysical and biochemical techniques and apply them together with site-directed spin labeling and multifrequency EPR spectroscopy, to explore internal dynamics of myosin motor functioning. The proposed work is based on recent developments by the applicant in sensitivity improvement for EPR measurement, especially for experiments on myosin in solution, and in multifrequency EPR data analysis, to accurately characterize the dynamics of a proteinbound spin label. This approach will be used to obtain detailed information about the dynamic rearrangement of protein structural elements. These methods will be developed further, and used to define the kinetics of structural rearrangements of the relay helix and SH1 helix in myosin during the actomyosin ATPase cycle.