Functional Dynamics of the Myosin Molecular Motor

申请人： 申请人 Yuri E. Nesmelov 博士，是一位物理学家，其研究重点是 电磁场与物质的相互作用。 Nesmelov 博士建议学习和利用分子 生物物理和生物化学技术完善了用于研究的多学科方法 肌球蛋白功能动力学。职业发展/培训计划：该计划的主要要素是 通过指导培训获得先进的分子生物物理和生化技术。的 大部分培训将来自主要导师 David D. Thomas 博士，他是国际知名的 肌肉生物物理学领域的知名科学家，并且是 NIH 资助的培训项目的主任 肌肉研究。细胞和分子生物学方面的额外培训将由共同导师 Dr. 玛格丽特·提图斯 (Margaret Titus)，因其在肌球蛋白方面的研究而闻名。部分研究（W波段电子 顺磁共振 (EPR)）将由著名专家 Peter Fajer 博士合作进行 生物 EPR 和肌肉生物物理学。进一步职业和研究发展的结构化活动 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，以及与明尼苏达大学肌肉萎缩症中心的联系。研究 计划：该提案的总体目标是让 Nesmelov 博士学习分子生物物理和生物化学 技术并将其与定点自旋标记和多频 EPR 光谱一起应用， 探索肌球蛋白运动功能的内部动态。拟议的工作基于最近的发展 由申请人改进EPR测量的灵敏度，特别是肌球蛋白的实验 解决方案，并在多频率 EPR 数据分析中，准确表征蛋白质的动力学 绑定的自旋标签。该方法将用于获取有关动态的详细信息 蛋白质结构元件的重排。这些方法将得到进一步发展，并用于定义 肌动球蛋白在肌球蛋白中中继螺旋和SH1螺旋结构重排的动力学 ATP 酶循环。

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 protein- bound 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.