Purification and characterization of NADH oxidase from Serpulina (Treponema) hyodysenteriae

Stanton, Thad B.; Jensen, Neil S.

doi:10.1128/jb.175.10.2980-2987.1993

Cited by 58 publications

(45 citation statements)

References 43 publications

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“…8,11,18,28,29,33,34,[39][40][41] None of these potential virulence attributes is likely to directly cause the lesions demonstrated in the present studies. In addition, three genes from S. hyodysenteriae, tlyA, tlyB, and tlyC, have been identified, cloned, and sequenced.…”

Section: Discussionmentioning

confidence: 89%

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

Hutto

Wannemuehler

1999

Vet Pathol

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Abstract. Studies were carried out to compare the early morphologic changes in the cecal mucosa of mice either infected with Serpulina hyodysenteriae or exposed to the ␤-hemolysin of S. hyodysenteriae. Sixty-five 12-24-week-old C3H/HeOuJ mice were infected with S. hyodysenteriae by gastric intubation. Two mice were necropsied every hour for 30 hours following infection. S. hyodysenteriae was isolated from the cecal contents of each mouse at all time points. Macroscopic lesions were first apparent at 14 hours postinfection (PI), and light microscopic lesions were first apparent at 10 hours PI, earlier than has been previously reported. Ultrastructural changes, first evident at 6 hours PI, included disarray and loss of microvilli and terminal web, with dilatation of intercellular spaces. Luminal bacteria were translocated through epithelial cells to the lamina propria, where capillaries exhibited changes indicative of increased permeability. In another experiment, solutions containing between 2,500 and 25,000 hemolytic units of purified S. hyodysenteriae hemolysin were placed within the lumen of surgically closed murine ceca (n ϭ 10); ceca were collected for examination 3 hours following treatment. Ultrastructural changes consisted of loss of microvilli and terminal web and marked vacuolation and exfoliation of epithelial cells. Significant numbers of necrotic and apoptotic epithelial cells were present, and epithelial cells internalized moderate numbers of bacteria. The hemolysin of S. hyodysenteriae induces some of the same early ultrastructural changes in the cecal epithelium of mice as occur following infection with S. hyodysenteriae. Based on the observed bacterial translocation, luminal bacteria also appear to play a unique role in lesion development in this model.

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Section: Discussionmentioning

confidence: 89%

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

Hutto

Wannemuehler

1999

Vet Pathol

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“…The only two fully characterised, N-terminally sequenced, water-producing NADH oxidases, from Streptococcus fueculis and Serpulinu hyodyseriteriae, are tlavoproteins which contain a thiol group as part of their active site (Ross and Claiborne, 1992;Stanton and Jensen, 1993). It is postulated that these NADH oxidases provide a means for microaerophilic organisms to contend with or utilise oxygen in their environment, perhaps by acting as a cytochrome-independent terminal oxidase replacing the mitochondrial cytochrome oxidase (Stanton and Jensen, 1993 ;Brown et al, 1995).…”

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confidence: 99%

“…NADH oxidases catalyse, without intermediate electron carriers, either a one-, two-or four-electron reduction of dioxygen, producing 0;-, H,O, or H,O respectively, and have been detected in bacteria (Hoskins et al, 1962;Reinards et al, 1981 ;Kliiinkes et al, 1985;Saeki et al, 1985;Koike et al, 1985;Schmidt et al, 1986;Cocco et al, 1988;Maeda et al, 1992;Park et al, 1992;Stanton and Jensen, 1993) and in the microaerophilic protozoa, Trichomonas vaginalis and Entunioebu histolytica (Linstead and Bradley, 1988 ;Lo and Reeves, 1980). Interestingly, water-pro-ducing NADH oxidases have been identified in microaerophilic organisms which contain a fermentative metabolism, lack functional haem groups, cytochromes, at least one of the conventional detoxification enzymes and glutathione (Hoskins et al, 1962;Lo and Reeves, 1980;Linstead and Bradley, 1988;Ross and Claiborne, 1992;Stanton and Jensen, 1993). The only two fully characterised, N-terminally sequenced, water-producing NADH oxidases, from Streptococcus fueculis and Serpulinu hyodyseriteriae, are tlavoproteins which contain a thiol group as part of their active site (Ross and Claiborne, 1992;Stanton and Jensen, 1993).…”

mentioning

confidence: 99%

“…Interestingly, water-pro-ducing NADH oxidases have been identified in microaerophilic organisms which contain a fermentative metabolism, lack functional haem groups, cytochromes, at least one of the conventional detoxification enzymes and glutathione (Hoskins et al, 1962;Lo and Reeves, 1980;Linstead and Bradley, 1988;Ross and Claiborne, 1992;Stanton and Jensen, 1993). The only two fully characterised, N-terminally sequenced, water-producing NADH oxidases, from Streptococcus fueculis and Serpulinu hyodyseriteriae, are tlavoproteins which contain a thiol group as part of their active site (Ross and Claiborne, 1992;Stanton and Jensen, 1993). It is postulated that these NADH oxidases provide a means for microaerophilic organisms to contend with or utilise oxygen in their environment, perhaps by acting as a cytochrome-independent terminal oxidase replacing the mitochondrial cytochrome oxidase (Stanton and Jensen, 1993 ;Brown et al, 1995).…”

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confidence: 99%

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A H₂O‐Producing NADH Oxidase from the Protozoan Parasite Giardia Duodenalis

Brown

Upcroft

1996

European Journal of Biochemistry

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We describe the purification of a H,O-producing NADH oxidase from the protozoan parasite Giardiu duodenalis. The enzyme is a monomeric flavoprotein containing flavin adenine dinucleotide in a 1 : 1 molar ratio with the polypeptide. The NADH oxidase has an apparent molecular mass of 46 kDa and was homogenous as determined by denaturing gel electrophoresis and N-terminal amino acid sequencing. NADPH could substitute for NADH as an electron donor with a K, value of 4.2 pM for NADH and 16 pM for NADPH (pH 7.8 at room temperature). With oxygen as the primary electron acceptor under aerobic conditions, the pure enzyme did not produce 0;-nor H,O, as stoichiometric products of oxygen reduction, implicating H,O as the end product and obviating the need for superoxide dismutase. The ability to utilise oxygen explains the apparent respiration of the amitochondrial fermentative metabolism of Giardia. Mercurials, flavoantagonists and heavy metals (CU'+ and ZnZ+ ) inhibited this activity. Under anaerobic conditions the enzyme catalysed electron transfer at lower efficiencies to other electron acceptors including nitroblue tetrazolium, potassium ferricyanide, FAD and FMN, using either NADH or NADPH as electron donors. NADPH, however, was a more efficient electron donor. Cytochrome c was not reduced under any assay conditions used. The enzyme reduced the nitrofuran drugs, furazolidone (an antigiardial) and nitrofurantoin, to their toxic radical forms as determined by EPR. Metronidazole, a nitroimidazole, was not reduced. Pure NADH oxidase did not demonstrate ferredoxin:NAD(P) ' oxidoreductase activity since it could not accept electrons from reduced ferredoxin to regenerate NAD(P)H. The G. duodenalis NADH oxidase may, therefore, function as a terminal oxidase, similar to the mitochondria1 cytochrome oxidase, and in the maintenance of an optimum intracellular redox ratio. This report of a flavoenzyme from Giardia places Giardiu close to the anaerobic bacteria in evolutionary terms.

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“…Catalase, peroxidase, NADH oxidase and superoxide dismutase activities were assayed according to standard protocols (Beauchamp & Fridovich, 1971;Beers & Sizers, 1952;Stanton & Jensen, 1993;Stellmach et al, 1988), and units for these activities are, respectively: 1 mmol H 2 O 2 consumed min…”

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confidence: 99%

Trophic interaction of the aerotolerant anaerobe Clostridium intestinale and the acetogen Sporomusa rhizae sp. nov. isolated from roots of the black needlerush Juncus roemerianus

et al. 2006

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Acetogens were enumerated from root homogenates of the black needlerush Juncus roemerianus obtained from a nearly pristine salt marsh. An isolated colony, ST1, yielded acetogenic activity and was initially thought to be a pure culture; however, ST1 was subsequently found to be composed of an aerotolerant fermentative anaerobe (RC) and an acetogen (RST) (T indicates type strain). The two spore-forming mesophiles were separated by selective cultivation under conditions favouring the growth of either RC or RST. The 16S rRNA gene sequence of RC was 99 % similar to that of Clostridium intestinale, indicating that RC was a new isolate of this clostridial species. The rRNA gene sequence most similar to that of RST was only 96 % similar to that of RST and was from a species of the acetogenic genus Sporomusa, indicating that RST was a new sporomusal species; the name Sporomusa rhizae sp. nov. is proposed. RC grew at the expense of saccharides. H2-forming butyrate fermentation was the primary catabolism utilized by RC under anoxic conditions, while homolactate fermentation was the primary catabolism under oxic conditions. RC consumed O2 and tolerated 20 % O2 in the headspace of shaken broth cultures. In contrast, RST was acetogenic, utilized H2, lactate and formate, did not utilize saccharides, and could not tolerate high concentrations of O2. RST grew by trophic interaction with RC on saccharides via the uptake of H2, and, to a lesser extent, lactate and formate produced by RC. Co-cultures of the two organisms yielded high amounts of acetate. These results indicate that (i) previously uncharacterized species of Sporomusa are associated with Juncus roots and (ii) trophic links to O2-consuming aerotolerant anaerobes might contribute to the in situ activities and survival strategies of acetogens in salt marsh rhizospheres, a habitat subject to gradients of plant-derived O2.

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Purification and characterization of NADH oxidase from Serpulina (Treponema) hyodysenteriae

Cited by 58 publications

References 43 publications

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

A H₂O‐Producing NADH Oxidase from the Protozoan Parasite Giardia Duodenalis

Trophic interaction of the aerotolerant anaerobe Clostridium intestinale and the acetogen Sporomusa rhizae sp. nov. isolated from roots of the black needlerush Juncus roemerianus

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Purification and characterization of NADH oxidase from Serpulina (Treponema) hyodysenteriae

Cited by 58 publications

References 43 publications

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

A Comparison of the Morphologic Effects ofSerpulina hyodysenteriaeor its Beta-hemolysin on the Murine Cecal Mucosa

A H2O‐Producing NADH Oxidase from the Protozoan Parasite Giardia Duodenalis

Trophic interaction of the aerotolerant anaerobe Clostridium intestinale and the acetogen Sporomusa rhizae sp. nov. isolated from roots of the black needlerush Juncus roemerianus

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A H₂O‐Producing NADH Oxidase from the Protozoan Parasite Giardia Duodenalis