Antigenic and Structural Analysis of Treponema denticola

Cockayne, Alan; Sanger, Ruth; Ivic, A.; Strugnell, R A; MacDougall, Jane; Russell, R; Penn, Charles W.

doi:10.1099/00221287-135-12-3209

Cited by 28 publications

(45 citation statements)

References 31 publications

(19 reference statements)

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“…PFs were com- prised of two major bands at 38 and 35 kDa and two minor bands at 32 and 29 kDa (42,43). These results are similar to those reported by Cockayne et al (9). Two-dimensional gel electrophoresis of whole-cell lysates (Fig.…”

Section: High-voltage Analysis Of Wild-type Cellssupporting

confidence: 82%

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

et al. 1997

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Treponema denticola is an anaerobic, motile, oral spirochete associated with periodontal disease. We found that the periplasmic flagella (PFs), which are located between the outer membrane sheath and cell cylinder, influence its morphology in a unique manner. In addition, the protein composition of the PFs was found to be quite complex and similar to those of other spirochetes. Dark-field microscopy revealed that most wild-type cells had an irregular twisted morphology, with both planar and helical regions, and a minority of cells had a regular right-handed helical shape. High-voltage electron microscopy indicated that the PFs, especially in those regions of the cell which were planar, wrapped around the cell body axis in a right-handed sense. In those regions of the cell which were helical or irregular, the PFs tended to lie along the cell axis. The PFs caused the cell to form the irregular shape, as two nonmotile, PF-deficient mutants (JR1 and HL51) were no longer irregular but were right-handed helices. JR1 was isolated as a spontaneously occurring nonmotile mutant, and HL51 was isolated as a site-directed mutant in the flagellar hook gene flgE. Consistent with these results is the finding that wild-type cells with their outer membrane sheath removed were also right-handed helices similar in shape to JR1 and HL51. Purified PFs were analyzed by two-dimensional gel electrophoresis, and several protein species were identified. Western blot analysis using antisera to Treponema pallidum PF proteins along with N-terminal amino acid sequence analysis indicated T. denticola PFs are composed of one class A sheath protein of 38 kDa (FlaA) and three class B proteins of 35 kDa (FlaB1 and FlaB2) and one of 34 kDa (FlaB3). The N-terminal amino acid sequences of the FlaA and FlaB proteins of T. denticola were most similar to those of T. pallidum and Treponema phagedenis. Because these proteins were present in markedly reduced amounts or were absent in HL51, PF synthesis is likely to be regulated in a hierarchy similar to that found for flagellar synthesis in other bacteria.Spirochetes are recognized for their unique cell morphology and unusual means of motility (5, 48). In most spirochetes, the primary structural component of the cell body is a flexible yet semirigid helically shaped cell cylinder (5). Borrelia burgdorferi is an exception, since it has a rod-shaped cell cylinder (13, 15). Helical periplasmic flagella (PFs) are closely associated with the spirochetal protoplasmic cell cylinder (PC) and are embedded in the cytoplasmic membrane and cell wall near each end of the cell (5, 6, 17); each PF is inserted at only one end. Surrounding the entire cell is an outer membrane sheath (OS) (5). Genetic evidence indicates that the PFs are essential for the motility of several spirochete species (4,8,26,28,37,44). The PFs vary in number, length, and protein composition among the spirochete species. Depending on the species, the PFs may or may not overlap in the center of the cell (5, 8). PFs from most spirochete species are co...

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Section: High-voltage Analysis Of Wild-type Cellssupporting

confidence: 82%

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

et al. 1997

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“…MgCl 2 ) induced a separation in aggregable and nonaggregable moieties, which could have enhanced minor parts of very large and perhaps different lipid molecules (such as the enterobacterial common antigen present in Enterobacteriaceae). Others suggest that an LPS-like structure exists in a different strain of T. denticola (ATCC 33520) on account of their findings of proteinase-indigestible low molecular weight molecules in silverstained gels (36). An interesting observation in our study is the actual size of OML521, which is around the same size as RaLPS, a structure essential to the assembly of perfectly active porin-trimers in normal outer membranes (37).…”

Section: Resultssupporting

confidence: 59%

Evidence for a New Type of Outer Membrane Lipid in Oral Spirochete Treponema denticola

Schultz¹,

Wolf²,

Lange³

et al. 1998

Journal of Biological Chemistry

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A new class of outer membrane lipid (OML) was isolated from the oral spirochete Treponema denticola strain ATCC 33521 using a phenol/chloroform/light petroleum procedure normally applied for lipopolysaccharide extraction. In addition to chemical analysis, Fourier transform infrared (FTIR) spectroscopy was applied to compare the biophysical properties of OML with lipopolysaccharides (LPS) and lipoteichoic acids (LTA). Isolated OML fractions represent 1.4% of the total dry cell weight, are about 4 kDa in size, and contain 6% amino sugars, 8% neutral sugars, 14% phosphate, 35% carbazol-positive compounds, and 11% fatty acids (containing iso-and anteiso-fatty acyl chains). Rare for outer membrane lipids, OML contains no significant amount of 3-deoxy-D-manno-octulosonic acids, heptoses, and ␤-hydroxy fatty acids. The fatty acyl chain composition, being similar to that of the cytoplasmic membrane, is quite heterogeneous with anteiso-pentadecanoic acid (12%), palmitic acid (51%), and iso-palmitic acid (19%) as the predominant fatty acids present. Findings of a glycerol-hexose unit and two glycerol-hexadecanoic acid fragments indicate a glycolipid membrane anchor typically found in LTA. There was also no evidence for the presence of a sphingosine-based lipid structure. The results of FTIR measurements strongly suggest that the reconstituted lipid forms normal bilayer structures (vesicles) expressing a high membrane state of order with a distinct phase transition as typical for isolated LPS. However, in contrast to LPS, OML of T. denticola has a lower T m near 22°C and a lower cooperativity of the phase transition. The results suggest a different kind of permeation barrier that is built up by this particular OML of T. denticola, which is quite different from LPS normally essential for Gram-negative bacteria.

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“…However, the mol. wt of 56-58 kDa for the OM protein reported here is different from that reported by Cockayne et al 42 and Cockayne et al 42 reported that four major OM polypeptides, of 73, 68, 54 and 52 kDa, were isolated from T. denticola ATCC 33520 by a Triton XlOO solubilisation procedure, whereas Yotis et al 43 reported that, for three strains of T. denticola, OM preparations extracted by SDS contained polypeptides of 66,53 and 45 kDa. Although the 53-kDa protein described by these authors may be a serovar-specific antigen, our results showed that the 5658-kDa protein was speciesspecific.…”

Section: Discussioncontrasting

confidence: 56%

“…However, without a comparative study involving all the isolation procedures used by the different groups, it is difficult to ascertain whether these proteins are truly one and the same. Cockayne et al 42 noted that the 54-kDa protein may be a breakdown product of a larger, heat-modifiable polypeptide. Other finding^^^.^^ suggest that the OM proteins may be involved in the binding of host serum proteins such as laminin, fibronectin and fibrinogen, or in the adherence to host tissues.…”

Section: Discussionmentioning

confidence: 99%

Outer membrane antigens of oral Treponema species

Tall

Nauman²

1994

Journal of Medical Microbiology

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Summary. Outer membrane (OM) antigens of several oral treponemes were studied. SDS-PAGE revealed a 56-58-kDa protein as a major component of isolated OM vesicles. Immunoblot analysis with eluted antibodies prepared from Treponema denticola ATCC 33520 recognised the 56-58-kDa protein, which was highly conserved in the 17 T. denticola strains tested. The protein was also a component of a T. denticola desoxycholate-extractable, ethanol-soluble antigen (DES-Ag) but was not present in T. pectinovorum or T. vincentii ATCC 35580 and strain N9 whole-cell lysates. Electronmicroscopy of OM vesicles showed typical treponemal ultrastructure. Immunogold labelling of T. denticola ATCC 33520 with T. denticola ATCC 3352Cspecific eluted antibodies recognised only the OM surface of the cell. These results suggest that the 56-58-kDa antigen comprises surface-orientated epitopes and that this antigen may be specific for T. denticola.

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Antigenic and Structural Analysis of Treponema denticola

Cited by 28 publications

References 31 publications

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

Evidence for a New Type of Outer Membrane Lipid in Oral Spirochete Treponema denticola

Outer membrane antigens of oral Treponema species

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