Darst S A, Ribi H O, Pierce D W, Kornberg R D
Department of Cell Biology, Stanford University School of Medicine CA 94305.
J Mol Biol. 1988 Sep 5;203(1):269-73. doi: 10.1016/0022-2836(88)90107-6.
Escherichia coli RNA polymerase holoenzyme forms two-dimensional crystals when adsorbed to positively charged lipid layers at the air/water interface. Adsorption of the protein is driven by electrostatic interactions between the positively charged lipid surface and the polymerase molecule, which has a net negative charge. Crystallization is dependent on the adsorption and concentration of RNA polymerase on fluid lipid surfaces. Image analysis of electron micrographs of crystals in negative stain, which diffract to 30 A resolution, shows irregularly shaped protein densities about 100 x 160 A, consistent with the dimensions of single polymerase molecules.
当大肠杆菌RNA聚合酶全酶吸附到空气/水界面带正电荷的脂质层上时,会形成二维晶体。蛋白质的吸附是由带正电荷的脂质表面与带净负电荷的聚合酶分子之间的静电相互作用驱动的。结晶取决于RNA聚合酶在流体脂质表面的吸附和浓度。对负染晶体的电子显微镜照片进行图像分析,这些晶体的衍射分辨率可达30埃,结果显示出不规则形状的蛋白质密度区域,大小约为100×160埃,与单个聚合酶分子的尺寸相符。