THE OMPC PROTEIN OF YERSINIA-ENTEROCOLITICA - PURIFICATION AND PROPERTIES
THE OMPC PROTEIN OF YERSINIA-ENTEROCOLITICA - PURIFICATION AND PROPERTIES
复制标题
DOI：
10.1016/0923-2508(89)90192-7
复制
发表时间：
1989-01-01
影响因子：
2.6
通讯作者：
BOOS, W
中科院分区：
文献类型：
--
作者：
BRZOSTEK, K;HREBENDA, J;BOOS, W
OmpC是小肠结肠炎耶尔森氏菌的主要外膜蛋白之一。当在室温下溶解时，这种蛋白质出现在SDS聚丙烯酰胺凝胶电泳作为一个寡聚体。加热至沸水温度后，单体的表观分子量为36，000。将纯化的OmpC掺入到黑色脂质膜中导致膜电导增加，表明孔形成活性。重构孔具有一般扩散孔的特征。它们显示出阳离子选择性，并且在1.0 M KCl中具有1.3 nS的单通道电导。假设孔的直径恒定，长度为6 cm（外膜的宽度），孔内外的离子电导率相同，则孔蛋白的直径估计为1.0 nm。多克隆抗体产生的天然，孔形成蛋白的制备。这些抗体不识别蛋白质的变性形式，但与大肠杆菌的天然OmpC和OmpF交叉反应。OmpC在Y.小肠结肠炎菌对渗透压的依赖性与R.杆菌
OmpC, one of the major outer membrane proteins of Yersinia enterocolitica, was isolated and purified to homogeneity. When solubilized at room temperature, this protein appeared on SDS polyacrylamide gel electrophoresis as an oligomer. After heating to the temperature of boiling water, the apparent molecular weight of the monomer was 36,000. The incorporation of purified OmpC into black lipid membranes resulted in an increase in membrane conductance demonstrating pore-forming activity. The reconstituted pores exhibited the charactertics of general diffusion pores. They showed cation selectivity and had a single channel conductance of 1.3 nS in 1.0 M KCI. Assuming a constant dimater of the pore, length of 6 cm (the width of the outer membrane) and the same ion conductivity inside and outside the pore, the diameter of the pore protein was estimated as 1.0 nm. Polyclonal antibodies were raised against the native, pore-forming protein preparation. These antibodies did not recognize the denatured form of the protein, but cross-reacted with native OmpC and OmpF of Escherichia coli. The reglation of OmpC expression in Y. enterocolitica was dependent on the osmolarity of the medium in the same way as in R. coli.