Watanabe Yasushi, Inoko Yoji
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Protein J. 2005 Apr;24(3):167-74. doi: 10.1007/s10930-005-7840-7.
The in vitro reassembled species of OmpF porin, which was renatured from its denatured monomer using n-octyl-beta-D-glucopyranoside, was characterized by low-angle laser light scattering photometry, circular dichroism spectroscopy and synchrotron radiation small-angle X-ray scattering measurements. The light scattering measurement reconfirmed that the reassembled species was the dimer of the protein. Circular dichroism spectra of the reassembled dimer showed a native-like beta-structure. A small-angle X-ray scattering measurement indicated that the size of the reassembled dimer was nearly equal to that of the native trimer under the present experimental conditions. In a thermal denaturation experiment followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the reassembled dimer was less stable than the native trimer.
使用正辛基-β-D-吡喃葡萄糖苷从变性单体复性得到的体外重新组装的OmpF孔蛋白物种，通过低角度激光光散射光度法、圆二色光谱法和同步辐射小角X射线散射测量进行了表征。光散射测量再次证实重新组装的物种是该蛋白质的二聚体。重新组装的二聚体的圆二色光谱显示出类似天然的β结构。小角X射线散射测量表明，在当前实验条件下，重新组装的二聚体的大小几乎与天然三聚体的大小相等。在随后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的热变性实验中，重新组装的二聚体比天然三聚体稳定性更低。
