Karch H, Leying H, Büscher K H, Kroll H P, Opferkuch W
Infect Immun. 1985 Feb;47(2):549-54. doi: 10.1128/iai.47.2.549-554.1985.
The fimbrial (pili) profile of a single strain of Escherichia coli O7:K1:H6 (WF96) was evaluated. Fimbriae were isolated by sucrose density gradient ultracentrifugation, purified from flagellae by the use of 0.4% sodium dodecyl sulfate (SDS), and separated into distinct fimbrial types. Analysis of the purified WF96 fimbriae by SDS-polyacrylamide gel electrophoresis revealed two polypeptide bands with molecular weights of 16,000 and 21,000. Treatment of the fimbrial mixture with saturated guanidine hydrochloride resulted in the appearance of a third band with a molecular weight of 19,500. The relative susceptibilities of the WF96 fimbrial types to disrupting chemicals (octyl-glucoside, urea, SDS, and guanidine hydrochloride) were assessed by exposure of the fimbrial mixture to each agent, separation of the depolymerized fimbriae from intact fimbriae by gel filtration on Sepharose CL-4B, and identification of the disaggregated fimbrial types by SDS-polyacrylamide gel electrophoresis of column fractions. The physicochemical heterogeneity of the three fimbrial types coexpressed on WF96 was exploited to develop a method for separation of individual fimbriae.
对一株大肠杆菌O7:K1:H6（WF96）的菌毛（菌毛）图谱进行了评估。通过蔗糖密度梯度超速离心法分离菌毛，使用0.4%十二烷基硫酸钠（SDS）从鞭毛中纯化菌毛，并将其分离为不同的菌毛类型。通过SDS-聚丙烯酰胺凝胶电泳对纯化的WF96菌毛进行分析，显示出两条分子量分别为16,000和21,000的多肽带。用饱和盐酸胍处理菌毛混合物后，出现了一条分子量为19,500的第三条带。通过将菌毛混合物暴露于每种试剂、通过Sepharose CL-4B凝胶过滤从完整菌毛中分离解聚的菌毛以及通过柱级分的SDS-聚丙烯酰胺凝胶电泳鉴定解聚的菌毛类型，评估了WF96菌毛类型对破坏化学物质（辛基葡糖苷、尿素、SDS和盐酸胍）的相对敏感性。利用在WF96上共表达的三种菌毛类型的物理化学异质性，开发了一种分离单个菌毛的方法。
