Excretion of flagellin by a short-flagella mutant of Salmonella typhimurium.
A nonmotile mutant of Salmonella typhimurium, SJW1254, has very short flagella (less than 0.1 micron long) due to a mutation in the structural gene of flagellin (H2). When ammonium sulfate was added to the culture medium of SJW1254 grown to the late-log phase, a large amount of protein precipitated. Gel electrophoresis and immunodiffusion showed that more than 90% (wt/wt) of the precipitated protein was flagellin. The mutant flagellin appeared to be excreted in the monomeric form, in an amount comparable to the amount in the flagellar filaments of wildtype bacteria. No such precipitate was obtained from the medium of wild-type bacteria. The mutant flagellin had the same apparent molecular weight (55,000) and isoelectric point (5.3) as the wild-type flagellin, but differed in mobility in polyacrylamide gel electrophoresis under nondenaturing conditions. Moreover, the mutant flagellin did not polymerize in vitro under various conditions in which wild-type flagellin polymerized. These results suggested that the mutant bacteria excreted flagellin because the flagellin polymerized poorly and therefore could not be trapped at the tip of the flagellar filament. This short-flagella mutant should be useful for studying the mechanism of flagellin transport.
D001426:Bacterial Proteins / Q000378:metabolism*; D005407:Flagella / Q000648:ultrastructure; D005408:Flagellin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007526:Isoelectric Point; D046911:Macromolecular Substances; D008970:Molecular Weight; D009154:Mutation; D012486:Salmonella typhimurium / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D001426:Bacterial Proteins; D046911:Macromolecular Substances; D005408:Flagellin