Schertler G F
MRC Laboratory of Molecular Biology, Cambridge, UK.
Eye (Lond). 1998;12 ( Pt 3b):504-10. doi: 10.1038/eye.1998.138.
Two-dimensional crystals of rhodopsin were studied to determine the arrangement of the transmembrane alpha helices. A combination of electron cryo-microscopy, image processing and electron crystallography was used to extract amplitudes and phases from images, and a three-dimensional map to a resolution of 7.5 A was calculated. Density peaks for all seven transmembrane helices were observed and the helix axes for all seven helices could be estimated. Near the intracellular side, which interacts with the G protein transducin, we observed three layers of helices arranged differently from bacteriorhodopsin. The arrangement opens up towards the extracellular side forming a cavity that serves as the binding pocket for the retinal. This cavity is closed towards the intracellular side by the long and highly tilted helix 3, and must be closed towards the extracellular side by the loop linking helices 4 and 5 that is linked by a disulphide bridge to the extracellular end of helix 3.
研究了视紫红质的二维晶体,以确定跨膜α螺旋的排列方式。采用电子冷冻显微镜、图像处理和电子晶体学相结合的方法,从图像中提取振幅和相位,并计算出分辨率为7.5埃的三维图谱。观察到所有七个跨膜螺旋的密度峰,并估计了所有七个螺旋的轴。在与G蛋白转导素相互作用的细胞内侧附近,我们观察到三层螺旋,其排列方式与细菌视紫红质不同。这种排列向细胞外侧开放,形成一个作为视黄醛结合口袋的腔。这个腔在细胞内侧被长而高度倾斜的螺旋3封闭,在细胞外侧必须被连接螺旋4和5的环封闭,该环通过二硫键与螺旋3的细胞外末端相连。