Ueno Y, Sato C
Computational Biology Research Center, National Institute of Advanced Industrial Science and Technology AIST, Tsukuba Central 6, Tsukuba, 305-8566 Japan.
Sci Prog. 2001;84(Pt 4):291-309. doi: 10.3184/003685001783238952.
Single particle analysis in electron microscopy allows direct observation of the reconstructed three-dimensional structures of protein molecules. This method enables a more comprehensive study of membrane proteins which have been problematic in structural studies using X-ray crystallography. These membrane proteins include the voltage-sensitive ion channel proteins, which play an important rule in neural activities, and have great medical significance. The method described is supported by the development of cryo-electron microscopy and the angular reconstitution method. This review summarizes certain principles governing single particle analysis employing angular reconstitution. This method was applied to our study of the voltage-sensitive sodium channel, and the results are discussed. With improvements in resolutions and statistical analyses, the single particle technique is considered to be advantageous in studies of the structural changes and molecular interactions of protein molecules.
电子显微镜中的单颗粒分析能够直接观察蛋白质分子重构后的三维结构。该方法有助于更全面地研究膜蛋白，而膜蛋白在利用X射线晶体学进行的结构研究中一直存在难题。这些膜蛋白包括电压敏感离子通道蛋白，它们在神经活动中起重要作用，具有重大医学意义。所述方法得到了冷冻电子显微镜和角度重构方法发展的支持。本综述总结了采用角度重构进行单颗粒分析的某些原理。该方法已应用于我们对电压敏感钠通道的研究，并对结果进行了讨论。随着分辨率和统计分析的改进，单颗粒技术在蛋白质分子结构变化和分子相互作用的研究中被认为具有优势。
