Purification and partial characterization of Bacillus subtilis Flagellar hooks.
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.
D000818:Animals; D000942:Antigens, Bacterial; D001412:Bacillus subtilis / Q000032:analysis / Q000166:cytology* / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification; D002499:Centrifugation, Density Gradient; D003168:Complement Fixation Tests; D003973:Diatrizoate; D004589:Electrophoresis, Disc; D005407:Flagella / Q000032:analysis / Q000276:immunology; D005827:Genetics, Microbial; D006358:Hot Temperature; D007106:Immune Sera; D008854:Microscopy, Electron; D009154:Mutation; D010772:Phosphotungstic Acid; D011489:Protein Denaturation; D011817:Rabbits; D012964:Sodium; D013194:Staining and Labeling; D013395:Sucrose; D013431:Sulfates
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D007106:Immune Sera; D013431:Sulfates; D003973:Diatrizoate; D010772:Phosphotungstic Acid; D013395:Sucrose; D012964:Sodium