Muir L L, Strugnell R A, Davies J K
Department of Microbiology, Monash University, Clayton, Victoria, Australia.
Infect Immun. 1988 Jul;56(7):1743-7. doi: 10.1128/iai.56.7.1743-1747.1988.
Pili of Neisseria gonorrhoeae are thought to be composed entirely of identical subunits, called pilin, that self-assemble in vitro. Previous pilus purification methods have relied on this latter point, and dissociation and reassociation of pilin subunits has yielded pilin preparations of high purity. Such a procedure could result in the loss of any pilus-associated proteins. We have developed a procedure for the isolation of intact native pili in a deoxycholate-urea buffer in which the pili are fractionated on the basis of size and hydrophobicity. Electron microscopy indicates that the pili are largely free from outer membrane vesicles and other cellular material. Electrophoretic analysis has shown that a number of proteins copurify with pilin. Antibodies to these proteins could be removed from an antiserum against whole piliated cells by absorption with piliated cells but not by absorption with nonpiliated cells. Hence, our results indicate that these proteins could be pilus associated.
淋病奈瑟菌的菌毛被认为完全由相同的亚基组成，这些亚基称为菌毛蛋白，能在体外自行组装。以往的菌毛纯化方法依赖于这一点，菌毛蛋白亚基的解离和重新结合产生了高纯度的菌毛蛋白制剂。这样的过程可能会导致任何与菌毛相关的蛋白质丢失。我们开发了一种在脱氧胆酸盐 - 尿素缓冲液中分离完整天然菌毛的方法，在该缓冲液中，菌毛根据大小和疏水性进行分级分离。电子显微镜显示菌毛基本不含外膜囊泡和其他细胞物质。电泳分析表明，有多种蛋白质与菌毛蛋白共同纯化。针对这些蛋白质的抗体可以通过用有菌毛细胞吸收从抗全菌毛细胞抗血清中去除，但不能通过用无菌毛细胞吸收去除。因此，我们的结果表明这些蛋白质可能与菌毛相关。
