Dimmitt K, Simon M I
J Bacteriol. 1971 Oct;108(1):282-6. doi: 10.1128/jb.108.1.282-286.1971.
A method for preparing bacterial flagellar hook structures is described. The method involves isolating intact flagella from a mutant which makes thermally labile flagellar filaments and heat-treating them to disaggregate the filament preferentially. The resulting hook preparation can be separated and purified by velocity and isopycnic centrifugation. The purified hooks sediment at a relative S value of 77. On acrylamide gel electrophoresis in sodium dodecyl sulfate, they show one major and a number of minor protein bands. The purified hooks can be used to immunize rabbits, and the resulting antiserum is hook-specific. These results support the notion that hooks are composed of a protein that differs from flagellin.
描述了一种制备细菌鞭毛钩结构的方法。该方法包括从产生热不稳定鞭毛丝的突变体中分离完整的鞭毛,并对其进行热处理以优先使鞭毛丝解聚。所得的钩制剂可通过速度离心和等密度离心进行分离和纯化。纯化的钩以相对S值77沉降。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳上,它们显示出一条主要蛋白带和一些次要蛋白带。纯化的钩可用于免疫兔子,所得抗血清具有钩特异性。这些结果支持了钩由不同于鞭毛蛋白的蛋白质组成这一观点。