Purification and Partial Characterization of<i>Bacillus subtilis</i>Flagellar Hooks

Dimmitt, K.

doi:10.1128/jb.108.1.282-286.1971

Cited by 35 publications

(13 citation statements)

References 12 publications

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“…Only two classes of mutants showed any flagellar-related structure. The hag mutants, in agreement with previous observations (1 5), had hook structures. The flaE mutants were found to have long curly The basal structures are intimately involved in both the regulation and function of…”

Section: Introductionsupporting

confidence: 92%

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The regulation of flagellar formation and function

Hilmen

Silverman

1974

J Supramol Struct

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The basal structure of the flagellum controls both activity and assembly. In order to define the steps involved in these processes, genetic analysis was performed. Twenty genes were found t o be required for the complete assembly and function of the organelle. FlaE controls the length of the hook, flaA is required both t o maintain flagellar structure and for chemotaxis, and flaI plays a role in regulating the synthesis of the entire structure. Mutations mapping close to flaI (the cfs mutations) release flagellar synthesis from control by catabolite repression.The basal structure was purified and isolated. On SDS acrylamide gel electrophoresis, it contained a t least six distinguishable components. One major band corresponded t o the hook subunit with an apparent molecular weight of 42,000 daltons. The others had apparent molecular weights of 60,000, 40,000, 28,000, 25,000, and 18,000 daltons. The genes that correspond t o these polypeptides have not been identified.In exploring the role of the mot and che genes, assays were developed for the function of individual flagellar filaments. The filaments were found t o rotate and rotation could be modulated by changing their direction. Chemotaxis results from the modulation of flagellar rotation. Using the rotation assay the response of nonmotile cells t o attractants and repellents was followed.

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Section: Introductionsupporting

confidence: 92%

“…(7). Furthermore by selective disaggregation the flagellar filament could be removed from intact B. subtilis flagella and the remaining hook and basal region isolated by velocity and isopycnic gradient centrifugation (5). The same approach has been used to purify the basal structures from E. coli.…”

Section: Isolation Of Flagellar Componentsmentioning

confidence: 99%

The regulation of flagellar formation and function

Hilmen

Silverman

1974

J Supramol Struct

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“…The HFC purification protocol allowed the isolation of 50 mg HFC protein and finally 0.2 nag of purified hook protein from the 5 1 culture. This is a much higher rate compared to other protocols for the isolation of the flagellar hook of other bacterial species [16][17][18][19].…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

Isolation and characterization of the flagellar hook of Campylobacter jejuni

Glenn-Calvo¹

1994

FEMS Microbiology Letters

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A method for purification of the flagellar hook of Campylobacter jejuni is described. The hook was shown to be composed of a subunit protein, which has a molecular mass of 92,000 and an isoelectric point of pI 4.8. A monoclonal antibody and a polyvalent antiserum was raised against the purified flagellar hook of C. jejuni. Immuno-electronmicroscopy revealed that the epitope recognized by the monoclonal antibody is surface-located. However, this antibody reacted only with the hook of the immunization strain, but not with other strains or other flagellated bacteria. Thus, our data indicate that the immunodominant epitopes are located on the surface of the hook and that these epitopes are strain-specific.

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“…The hook of bacterial flagella is the structure that connects the filament to the basal body (7,8). It has been presumed to act as a flexible universal joint for the rotational movement of the fllament driven by a rotary motor residing in the basal body (29).…”

mentioning

confidence: 99%

“…It has been presumed to act as a flexible universal joint for the rotational movement of the fllament driven by a rotary motor residing in the basal body (29). Structurally, it is a cylindrical polymer of a single kind of protein, which is chemically and immunologically different from flagellin, the component protein of the filament (8,15,16).…”

mentioning

confidence: 99%

Role of gene flaFV on flagellar hook formation in Salmonella typhimurium

Kutsukake¹,

Suzuki

Yamaguchi

et al. 1979

J Bacteriol

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Nine temperature-sensitive nonflagellate mutants defective in flaFV were isolated from a strain of Salmonella typhimurium. Among them, three mutants were found to produce flagella with abnormally shaped (either straight or irregularly curved) hooks at the permissive temperature. Two mutations that rendered hooks straight were located in one of the eight segments of flaFV defined by deletion mapping. The mutation that rendered hooks irregularly curved was located in a different segment. An flaR mutation was introduced into the latter mutant. At the permissive temperature, the resulting double mutant produced polyhooks whose wavelength and amplitude were both exceedingly reduced. These polyhook structures were more thermolabile than those of the flaFV+ strain. Hook protein of the former strain was shown to have a slightly positive electric charge compared with that of the latter. From these results and other available information, it is inferred that flaFV is the structural gene for the hook protein in Salmonella.

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Purification and Partial Characterization ofBacillus subtilisFlagellar Hooks

Cited by 35 publications

References 12 publications

The regulation of flagellar formation and function

The regulation of flagellar formation and function

Isolation and characterization of the flagellar hook of Campylobacter jejuni

Role of gene flaFV on flagellar hook formation in Salmonella typhimurium

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