Hustedt E J, Beth A H
Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee 37232, USA.
Annu Rev Biophys Biomol Struct. 1999;28:129-53. doi: 10.1146/annurev.biophys.28.1.129.
Measurement of the distance between two spin label probes in proteins permits the spatial orientation of elements of defined secondary structure. By using site-directed spin labeling, it is possible to determine multiple distance constraints and thereby build tertiary and quaternary structural models as well as measure the kinetics of structural changes. New analytical methods for determining interprobe distances and relative orientations for uniquely oriented spin labels have been developed using global analysis of multifrequency electron paramagnetic resonance data. New methods have also been developed for determining interprobe distances for randomly oriented spin labels. These methods are being applied to a wide range of structural problems, including peptides, soluble proteins, and membrane proteins, that are not readily characterized by other structural techniques.
测量蛋白质中两个自旋标记探针之间的距离,可以确定特定二级结构元件的空间取向。通过使用定点自旋标记,能够确定多个距离限制,从而构建三级和四级结构模型,并测量结构变化的动力学。利用多频电子顺磁共振数据的全局分析,已开发出用于确定唯一取向自旋标记的探针间距离和相对取向的新分析方法。还开发了用于确定随机取向自旋标记的探针间距离的新方法。这些方法正被应用于广泛的结构问题,包括肽、可溶性蛋白质和膜蛋白,而这些用其他结构技术难以表征。