Stanislavsky E S, Edvabnaya L S, Zaidher I G, Makarenko T A, Bulk V F, Zhvanetskaya M I, Mashilova G M
Acta Microbiol Hung. 1986;33(3):245-55.
Crude aqueous extract was obtained from acetone-dried cells of Pseudomonas aeruginosa strain 868 (serogroup O2, Lányi and Bergan's schema) and subjected to ultracentrifugation (105,000 g, 3 h); the lipopolysaccharide (LPS)-containing precipitate was discarded and the supernatant containing water-soluble cell proteins was subjected to further fractionation. From a partially purified aqueous extract two fractions were obtained by step-wise precipitation with ammonium sulphate, namely, F1 (by 50% saturation), and F2 (by 80% saturation). By gel and ion-exchange chromatography from both fractions 9 subfractions were isolated differing in molecular weight, protein content, and LPS contamination. Subfractions 4 and 7 were practically free from LPS, and gave one precipitation line with antisera for strain 868. By immunoelectrophoresis subfraction 4 contained 2 cathodic and 1 anodic, whereas subfraction 7 mainly 1 anodic component. These subfractions were antigenically identical. With ELISA these subfractions were less active as compared to other subfractions, in particular to those of high molecular weight. The anti-subfraction 4 and anti-subfraction 7 sera were found to protect passively mice against intraperitoneal challenge by P. aeruginosa strain 8 (serogroup O2). These data support the authors' opinion that subfraction SF-4 and SF-7 are protective protein antigens (mol wt about 40,000 and 30,000, respectively), that are localized in the outer membrane of P. aeruginosa cell envelope.
从铜绿假单胞菌868菌株（O2血清群，Lányi和Bergan分类法）的丙酮干燥细胞中获得粗水提取物，并进行超速离心（105,000 g，3小时）；弃去含脂多糖（LPS）的沉淀，将含有水溶性细胞蛋白的上清液进行进一步分级分离。通过用硫酸铵逐步沉淀从部分纯化的水提取物中获得两个级分，即F1（50%饱和度）和F2（80%饱和度）。通过凝胶和离子交换色谱从这两个级分中分离出9个亚级分，它们在分子量、蛋白质含量和LPS污染方面有所不同。亚级分4和7几乎不含LPS，与868菌株抗血清产生一条沉淀线。通过免疫电泳，亚级分4含有2个阴极成分和1个阳极成分，而亚级分7主要含有1个阳极成分。这些亚级分在抗原性上是相同的。与ELISA相比，这些亚级分的活性较低，特别是与那些高分子量的亚级分相比。发现抗亚级分4和抗亚级分7血清能被动保护小鼠免受铜绿假单胞菌8菌株（O2血清群）的腹腔攻击。这些数据支持了作者的观点，即亚级分SF - 4和SF - 7是保护性蛋白抗原（分子量分别约为四万和三万），它们位于铜绿假单胞菌细胞包膜的外膜中。
