Hollingdale M R, Lemcke R M
J Hyg (Lond). 1972 Mar;70(1):85-98. doi: 10.1017/s0022172400022129.
Extraction of membranes of Mycoplasma hominis with n-butanol showed that antigenicity was associated with the non-lipid residue, which probably consisted mainly of protein, and not with the lipid itself.Since many membrane proteins are hydrophobic, membranes were rendered soluble in various ways. Extraction with urea or phenol was the most successful, yielding extracts which were both antigenic and serologically reactive. The urea extract could not be fractionated by polyacrylamide disk electrophoresis or by column chromatography. However, serologically active components identified by gel diffusion were separated from detergent-lysed membranes by polyacrylamide disk electrophoresis. The activities of antisera against these fractions suggested that indirect-haemagglutinating or metabolic-inhibiting antibodies can be directed against several different membrane antigens. However, the antigens identified by gel diffusion probably do not represent all the components participating in indirect haemagglutination.Treatment of membrane suspensions with heat, alkali, periodate and various enzymes showed that the four components identified by gel diffusion could be distinguished by their differing stabilities and properties. On the basis of their lability and susceptibility to proteolytic enzymes, two were identified as proteins.
用人型支原体膜与正丁醇进行提取,结果表明抗原性与非脂质残余物相关,该残余物可能主要由蛋白质组成,而非与脂质本身相关。由于许多膜蛋白具有疏水性,因此通过各种方式使膜变得可溶。用尿素或苯酚提取最为成功,得到的提取物既具有抗原性又具有血清学反应活性。尿素提取物不能通过聚丙烯酰胺圆盘电泳或柱色谱法进行分级分离。然而,通过凝胶扩散鉴定的具有血清学活性的成分通过聚丙烯酰胺圆盘电泳从去污剂裂解的膜中分离出来。针对这些组分的抗血清活性表明,间接血凝或代谢抑制抗体可针对几种不同的膜抗原。然而,通过凝胶扩散鉴定的抗原可能并不代表参与间接血凝的所有成分。用热、碱、高碘酸盐和各种酶处理膜悬浮液表明,通过凝胶扩散鉴定的四种成分可因其不同的稳定性和性质而区分。根据它们对蛋白水解酶的不稳定性和敏感性,其中两种被鉴定为蛋白质。