EPR Spectrometer Upgrade for Structural Biophysics Core Laboratory

明尼苏达大学的四个主要用户和五个次要用户正在获得EPR仪器的升级，以研究参与肌肉收缩、膜钙运输、光合作用、脂结合和运输以及蛋白质折叠的蛋白质的动态、结构和功能。研究人员正在利用最近成立的结构生物物理核心实验室(SBCL)的共用设施，通过诱变、蛋白质设计、生理学和酶动力学以及各种结构技术来研究生物功能和机制。新仪器具有以下功能：(1)可以对小样本和贵重样品进行实验，如突变蛋白质、单肌肉纤维和单蛋白晶体，并能够为时间分辨EPR实验提供可编程数据采集；(2)包括用于小取向样品和时间分辨光化学EPR实验的专用谐振器，以及(3)具有第二高的光谱仪频率，以提高数据分析的灵敏度和分辨率，并减少数据分析的模糊性。EPR可以同时检测肌肉结构蛋白的结构和动力学，并与肌肉的老化过程相关联。瞬变和多频电子顺磁共振将被用来探索光系统II--光合作用的水氧化复合体--催化部位的周转动力学。用溶液和单晶中的EPR研究脂结合蛋白和结合脂之间的动态相互作用，并与X射线结晶学相配合。

9970177SummaryFour major and five minor users at the University of Minnesota are acquiring an EPRinstrumentation upgrade to study the dynamics, structure, and function of proteins involved in muscle contraction, membrane calcium transport, photosynthesis, lipid binding and transport, and protein folding. The investigators are studying biological function and mechanism through mutagenesis, protein design, physiology, and enzyme kinetics, as well as a variety of structural techniques, using the shared facilities of a recently established Structural Biophysics Core Laboratory (SBCL). The new instrument has capabilities that will (1) allow experiments on small and precious samples, such as mutant proteins, single muscle fibers, and single protein crystals, and the capability for programmable data-acquisition for time-resolved EPR experiments, (2) include specialized resonators for small oriented samples and for time-resolved photochemical EPR experiments, and (3) has a second higher spectrometer frequency, to increase sensitivity and resolution and decrease ambiguity of data analysis. The structure and dynamics of muscle structural proteins will be detected by EPR simultaneously with muscle mechanics and correlated with the aging process. Transient and multi-frequency EPR will be used to probe the kinetics of turnover of the catalytic site of photosystem II, the photosynthetic water-oxidizing complex. The dynamic interaction between lipid binding proteins and bound lipids will be probed by EPR in solution and single protein crystals and coordinated with x-ray crystallography.