The Nature of Flagellar Antigens

Koffler, Henry; Smith, R. W.

doi:10.1007/978-1-4612-9843-4_4

Cited by 3 publications

(4 citation statements)

References 88 publications

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“…Our immunoprecipitation results suggest that the presence of multiple PF protein species is not due to phase variation or to different types of PFs residing on a given cell. The results presented here indicate that both proteins reside on a given PF in T. phagedenis, in analogy to C. crescentus (26,41) and B. pumilis (24,37).…”

Section: Discussionmentioning

confidence: 60%

“…Third, there could be one type of PF, with each PF consisting of multiple protein species. Both Bacillus pumilis (24,37) and Caulobacter crescentus (26,41) have multiple protein species associated on a given flagellum. We present evidence in this paper in support of the third explanation for T. phagedenis, i.e., there is more than one protein species residing on a given PF in T. phagedenis.…”

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confidence: 99%

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Treponema phagedenis has at least two proteins residing together on its periplasmic flagella

Limberger¹,

Charon²

1986

J Bacteriol

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Treponema phagedenis is an anaerobic, motile spirochete with several periplasmic flagella (PFs) at each cell end. This study provides the first genetic evidence that multiple protein species are associated with the PFs. In addition, these proteins were found to reside together on a given PF. Nonmotile mutants which lacked the PFs were isolated, and spontaneous revertantsWto motility regained the PFs. These results suggest that the PFs are involved in the motility of T. phagedenis. Isolated PFs had two major protein bands with molecular weights of 33,000 and 39,800, as revealed by sodium dodecyl sulfate-polyacrylamide, gel electrophoresis. Western blots with monoclonal and polyclonal antibodies indicated that both proteins were absent in the PF mutants but present in the revertants. Immunoelectron microscopy revealed that the 39,800-molecular-weight protein was distributed along the entire PF. Immunoprecipitation analysis suggested that the 39,800-and 33,000-molecular-weight proteins were closely associated in situ.Treponema phagedenis is an anaerobic spirochete that is cultivable on enriched media (36). The cells are approximately 0.2 ,um wide and consist of a right-handed helical cell cylinder (23) surrounded by an outer membrane sheath (9, 17-21, 31, 36, 38). Inserted subterminally at each end of the cell, between the helical cell cylinder and the outer membrane sheath, are several periplasmic flagella (PFs), also referred to as axial filaments, endoflagella, and periplasmic fibrils (9, 17-20, 31, 36, 38).The PFs of various spirochetes have been analyzed in some detail. Whereas most bacterial flagella consist of a single major protein, the major proteins isolated from PFs vary from one to six (5,6,16,22,28,30,32). In these studies, the major criterion used for the identification of PF proteins was enrichment during purification. In this paper, we present the first genetic evidence that multiple protein species are indeed associated with PFs.The finding of multiple protein species associated with PFs is consistent with three possibilities. Firsi, there could be two types of PFs, with each type consisting of a different protein. Both types of PFs could be found on each cell. This situation is analogous to the morphological flagellar types found in Vibrio parahaemolyticus (33). Second, phase variation could be occurring in the cell population. Each cell could be synthesizing only one or the other PF type. Salmonella species undergo such a phase variation in their flagella (34). Third, there could be one type of PF, with each PF consisting of multiple protein species. Both Bacillus pumilis (24, 37) and Caulobacter crescentus (26, 41) have multiple protein species associated on a given flagellum. We present evidence in this paper in support of the third explanation for T. phagedenis, i.e., there is more than one protein species residing on a given PF in T. phagedenis.( ). Cells were plated on PYG-RS plus 0.5% Bacto-Agar (Difco Laboratories, Detroit, Mich.) by using overlays made with 0.5% SeaPlaque agarose (FMC C...

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Section: Discussionmentioning

confidence: 60%

mentioning

confidence: 99%

Treponema phagedenis has at least two proteins residing together on its periplasmic flagella

Limberger¹,

Charon²

1986

J Bacteriol

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“…The absence of detectable cysteine in antigen [a] is a characteristic shared by other bacterial proteins which are secreted from cells (33) or which form structural components of the cell external to the cell wall proper. Its overall chemical composition resembles not only the outer protein of S. serpens, but other bacterial proteins, including flagellins (7,16,19), pilus proteins (21), colicin K (15), and the surface protein of Acinetobacter (40). It has been proposed that the deficiency in disulfide bonds common to such proteins is an essential feature of proteins which must pass through the rigid cell wall (33) and accounts for the ease with which they undergo conformational transformations (5).…”

Section: Discussionmentioning

confidence: 99%

Microcapsule of Campylobacter fetus: chemical and physical characterization

Winter¹,

McCoy²,

Fullmer³

et al. 1978

Infect Immun

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The antiphagocytic antigen (antigen [a]) comprising the microcapsule of a strain of Campylobacter fetus subsp. intestinalis has been purified from culture supernatants by ammonium sulfate fractionation and free-flow electrophoresis. Antigen [a] is a glycoprotein containing about 4% carbohydrate consisting of hexose, pentose, and methylpentose. The composition of the protein was typical of bacterial extramural structural proteins in its low content of basic, aromatic, and sulfur-containing amino acids. The protein had a high content of aspartic acid, threonine, glycine, and alanine. Antigen [a] had an Rf of 0.33 on polyacrylamide gel electrophoresis and a molecular weight calculated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of approximately 98,000. In contrast to its free form in culture supernatants, antigen [a] in vesicles derived from sheared cells appeared to exist in a complex with lipopolysaccharide. This complex could be dissociated by ethylenediaminetetraacetic acid or by ethylenediaminetetraacetic acid plus Triton X-100. A mutant strain that lacked a microcapsule, when incubated with soluble antigen [a] in a calcium medium, became agglutinable by monospecific [a] antiserum and showed an additional structural layer similar in appearance to the microcapsule on its cell wall. Points of similarity are emphasized between antigen [a] of C. fetus and the outer structural protein of the taxonomically related Spirillum serpens. An antiphagocytic surface component, designated antigen [a], has been defined on a strain of Campylobacter fetus subsp. intestinalis (24). This structure corresponds to a microcapsule on the basis of its morphological and functional attributes (42). The present study was conducted to further characterize antigen [a] and, in particular, to compare it with the "outer structured layer" (4, 5) of the taxonomically related bacterium Spirillum serpens. MATERIALS AND METHODS Bacterial strains and cultivation. C. fetus subsp. intestinalis strains 23B and 23D were employed. Strain 23D is the wild type which possesses the antiphagocytic component antigen [a], whereas mutant strain 23B lacks this antigen (24). Cultivation of organisms on plates, in broth, or in bottle overlay cultures was performed by methods previously described (25). Purification of antigen [a]. Cells of strain 23D were separated by centrifugation after cultivation for 16 h in Albimi broth (Difco Laboratories, Detroit, Mich.). Proteins were precipitated from the cell-free supernatant by addition of solid (NH4)2SO4 (50 g/100

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“…The bacterial flagellum is composed of a least three antigenically distinct structural units: the filament, hook, and basal disks. The flagellar filament is a tubule varying in size from 12 to 15 nm, composed of identical protein subunits in one of two configurations (1,15). Lowy and Hanson (16,17) demonstrated that the arrangement of subunits in intact flagella gives rise to two different types of filament configuration: an A form, which is characterized by helically connected globules, and a thick-lined B form in which helical organization is not observed.…”

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Flagellar ultrastructure and flagella-associated antigens of Campylobacter fetus

McCoy¹,

Doyle²,

Wiltberger³

et al. 1975

J Bacteriol

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Ultrastructural examinations of the flagellum of Compylobacter (Vibrio) fetus were performed throughout the growth cycle. Filament diameters, exceeding 17.6 nm during the exponential phase, were substantially greater than those reported for unsheathed flagella of other genera with the exception of Pseudomonas fluorescens. Filament diameters increased during growth, reaching a mean width of 21.2 nm in middle to late stationary phase. Internal flagellar structure, principally of the parallel lined variety, was observed during the later periods of growth but not during exponential or early stationary phase. Despite the unusually large filament sizes, no evidence of a flagellar sheath was observed after selected treatments (0.01 N HCl, 6 M urea, tris(hydroxymethyl) amino-methane-hydrochloride buffer, warm water) or examination of thin sections. To determine whether alterations in filament size and variable ability to demonstrate filament fine structure were correlated with progressive changes in serological activity, agglutination and immobilization tests were conducted with antisera directed against intact flagella, the principal flagellar antigen, the O antigen, and a superficial glycoprotein which has been found in association with the flagellum and the cell envelope. Significant differences in the serological activity of cells at different growth intervals were not noted with any of the sera employed.

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The Nature of Flagellar Antigens

Cited by 3 publications

References 88 publications

Treponema phagedenis has at least two proteins residing together on its periplasmic flagella

Treponema phagedenis has at least two proteins residing together on its periplasmic flagella

Microcapsule of Campylobacter fetus: chemical and physical characterization

Flagellar ultrastructure and flagella-associated antigens of Campylobacter fetus

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