Purification and Partial Characterization of Bacillus subtilis Flagellar Hooks

Abstract

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.