Ogasawara M, Kobayashi S, Arai S, Laheji K, Hill J L, Kono D H, Yu D T
J Immunol. 1985 Aug;135(2):1430-6.
The outer membranes of gram-negative bacteria are considered to be of importance in host-bacteria interaction, in protective immunity, and occasionally in subclassification within a species. In this study, the outer membranes of several strains of Yersinia enterocolitica and Y. pseudotuberculosis were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). It was found that the appearance of the major proteins depended on the temperature at which they were solubilized in SDS. A protein was identified with the use of two-dimensional gels and preparative SDS-PAGE, which was equivalent to the "heat-modifiable protein" (protein II) of other Enterobacteriaceae species. A monoclonal antibody, 4G1, was generated against an isolated preparation of this Y. enterocolitica protein. This antibody was tested with whole cell bacterial antigens of 46 individual bacterial strains. The reactive strains included only Y. enterocolitica and Y. pseudotuberculosis. In addition, the reactivity of the 4G1 monoclonal antibody preparation could be absorbed only with Y. enterocolitica and Y. pseudotuberculosis, and not with other strains of bacteria. The reactivity of this 4G1 monoclonal antibody was also tested by the Western Blot technique. Six individual strains were tested: a Y. enterocolitica serotype 0:3, a Y. enterocolitica serotype 0:9, an Escherichia coli, a Salmonella typhimurium, a Shigella flexneri, and a Klebsiella pneumoniae. The 4G1 antibody reacted with only the proteins of the two Y. enterocolitica strains. In conclusion, the equivalent of the heat-modifiable protein was present in Y. enterocolitica and Y. pseudotuberculosis. Moreover, this protein also carried a species-specific antigenic determinant.
革兰氏阴性菌的外膜被认为在宿主与细菌的相互作用、保护性免疫以及偶尔在物种内的亚分类中具有重要意义。在本研究中，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析了几株小肠结肠炎耶尔森菌和假结核耶尔森菌的外膜。发现主要蛋白质的出现取决于它们在SDS中溶解时的温度。使用二维凝胶和制备性SDS-PAGE鉴定出一种蛋白质，它等同于其他肠杆菌科物种的“热修饰蛋白”（蛋白II）。针对这种小肠结肠炎耶尔森菌蛋白的分离制剂产生了一种单克隆抗体4G1。用46个单个菌株的全细胞细菌抗原对该抗体进行了检测。反应性菌株仅包括小肠结肠炎耶尔森菌和假结核耶尔森菌。此外，4G1单克隆抗体制剂的反应性仅能被小肠结肠炎耶尔森菌和假结核耶尔森菌吸收，而不能被其他细菌菌株吸收。还通过蛋白质印迹技术检测了这种4G1单克隆抗体的反应性。测试了六个单个菌株：一株小肠结肠炎耶尔森菌0:3血清型、一株小肠结肠炎耶尔森菌0:9血清型、一株大肠杆菌、一株鼠伤寒沙门氏菌、一株弗氏志贺菌和一株肺炎克雷伯菌。4G1抗体仅与两株小肠结肠炎耶尔森菌的蛋白质发生反应。总之，小肠结肠炎耶尔森菌和假结核耶尔森菌中存在等同于热修饰蛋白的物质。此外，这种蛋白质还带有物种特异性抗原决定簇。
