Tortorello M L, Dunny G M
J Bacteriol. 1985 Apr;162(1):131-7. doi: 10.1128/jb.162.1.131-137.1985.
High-molecular-weight surface antigens, obtained by ammonium sulfate precipitation of culture supernatants and identified in Western blots of sodium dodecyl sulfate-polyacrylamide gels, have been correlated with the sex pheromone response of Streptococcus faecalis donor cells. Pheromone-induced cells carrying the conjugative plasmid pCF10 produced both an antigenic component (C130) composed of at least four bands in the range of 130 kilodaltons and a 73-kilodalton antigen (SA 73). The concentration of the C130 antigen in culture supernatants increased with time after exposure of donor cells to pheromone preparations. Gel filtration studies indicated that this antigen exists in the native state as a very large complex that is more than 180,000 daltons in size. The C130 antigen was susceptible to digestion by proteinase K and was not reactive with either concanavalin A or wheat germ agglutinin. The antigenicity of C130 was not destroyed by treatment of blots with trypsin, chymotrypsin, or papain before development with antibody, whereas the antigenicity of SA73 was susceptible to these treatments.
通过硫酸铵沉淀培养上清液获得的高分子量表面抗原,经十二烷基硫酸钠-聚丙烯酰胺凝胶的蛋白质印迹法鉴定,已与粪肠球菌供体细胞的性信息素反应相关。携带接合质粒pCF10的信息素诱导细胞产生了一种抗原成分(C130),该成分由至少四条在130千道尔顿范围内的条带组成,以及一种73千道尔顿的抗原(SA 73)。供体细胞暴露于信息素制剂后,培养上清液中C130抗原的浓度随时间增加。凝胶过滤研究表明,这种抗原以天然状态存在,是一种大小超过180,000道尔顿的非常大的复合物。C130抗原易被蛋白酶K消化,且与伴刀豆球蛋白A或麦胚凝集素均无反应。在用抗体显色之前,用胰蛋白酶、胰凝乳蛋白酶或木瓜蛋白酶处理印迹不会破坏C130的抗原性,而SA73的抗原性对这些处理敏感。