Ostrovskiĭ D N, Urban K, Reshetniak V I, Solton M
Biokhimiia. 1981 Jul;46(7):1321-30.
Incubation of Micrococcus lysodeikticus protoplasts with rabbit immunoglobulins containing antibodies against micrococcal membranes and a subsequent two-dimensional immunoelectrophoresis of solubilized membrane antigens allowed to reveal four main antigens of the outer membrane, of which only two are accessible to trypsin attack. The outer antigens disappear after protoplast treatment with snail digestive juice (active substance - phospholipase) and after incubation of growing cells with cerulenin, the inhibitor of fatty acid biosynthesis. Electrophoresis of the "cerulenin" membrane proteins in the presence of SDS revealed the disappearance of five proteins with molecular weights of 120 000, 150 000, 100 000, 68 000 and 16 000. Goat antibodies against rabbit serum or Staphylococcus aureus Cowan I cells (binding IgG gamma-globulins for fragment Fc) being added to the solubilized micrococcal membranes from protoplasts pretreated with antimembrane rabbit gamma-globulins precipitate about 5% of membrane protein. This suggests that the membrane outer layer antigens make up to about 5% of total protein content. The distribution of the components between the outer and inner membrane layers is discussed.
将溶壁微球菌原生质体与含有抗微球菌膜抗体的兔免疫球蛋白一起温育,随后对溶解的膜抗原进行二维免疫电泳,从而揭示出外膜的四种主要抗原,其中只有两种抗原可被胰蛋白酶攻击。在用蜗牛消化液(活性物质 - 磷脂酶)处理原生质体后以及在用脂肪酸生物合成抑制剂浅蓝菌素培养生长细胞后,外抗原消失。在十二烷基硫酸钠存在下对“浅蓝菌素”膜蛋白进行电泳,结果显示分子量为120000、150000、100000、68000和16000的五种蛋白质消失。将抗兔血清的山羊抗体或金黄色葡萄球菌考恩I细胞(结合IgGγ - 球蛋白的Fc片段)添加到用抗膜兔γ - 球蛋白预处理的原生质体溶解的微球菌膜中,沉淀出约5%的膜蛋白。这表明膜外层抗原约占总蛋白含量的5%。文中还讨论了各成分在外膜层和内膜层之间的分布情况。