Rappoport Joshua Z, Simon Sandford M, Benmerah Alexandre
The Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, Box 304, New York, New York 10021, USA.
Traffic. 2004 May;5(5):327-37. doi: 10.1111/j.1398-9219.2004.00187.x.
Most knowledge of clathrin-mediated endocytosis has been gained by biochemical fractionation and in vitro assays. Recently, the study of endocytosis has extended into the living cell. The tracking of individual clathrin-coated pits and vesicles (CCPs and CCVs) has provided new insight into understanding the dynamic nature of CCPs. The use of total internal reflection fluorescence microscopy (TIR-FM), also termed evanescent field microscopy, has enabled the direct observation of events occurring within a restricted area of the cell adjacent to and including the adherent plasma membrane. TIR-FM is now actively being pursued in the study of endocytic processes. The direct observation of CCP-associated proteins including clathrin itself, dynamin and, most recently, AP-2 has considerably challenged old models, confirming some points but raising very interesting new questions.
大多数关于网格蛋白介导的内吞作用的知识是通过生化分级分离和体外实验获得的。最近，内吞作用的研究已扩展到活细胞中。对单个网格蛋白包被小窝和囊泡（CCP和CCV）的追踪为理解CCP的动态性质提供了新的见解。全内反射荧光显微镜（TIR-FM）的使用，也称为倏逝场显微镜，使得能够直接观察在与附着质膜相邻并包括附着质膜的细胞受限区域内发生的事件。TIR-FM目前正积极应用于内吞过程的研究。对包括网格蛋白本身、发动蛋白以及最近的AP-2在内的与CCP相关蛋白的直接观察对旧模型提出了相当大的挑战，证实了一些观点，但也提出了非常有趣的新问题。
