Binding of hemin and Congo red by oral hemolytic spirochetes

Scott, D.; Siboo, Ian R.; Chan, E. C. S.; Klitorinos, A.; Siboo, R.

doi:10.1111/j.1399-302x.1993.tb00568.x

Cited by 22 publications

(27 citation statements)

References 35 publications

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“…Although CR-and/or haemin-binding phenotypes have been observed in a number of pathogenic organisms, including A. salmonicida (Kay et al, 1985), enteroinvasive E. coli (Stugard et al, 1989), Neisseria meningitidis (Payne & Finkelstein, 1977), Porphyromonas gingivalis (Genco et al, 1994), Shigella flexneri (Daskaleros & Payne, 1987;Stugard et al, 1989), Treponema species (Scott et al, 1993), Vibrio chlolerae (Payne & Finkelstein, 1977), Yersinia enterocolitica (Prpic et al, 1983) and highly variable pigmented Yersinia pseudotu6erculosis (Burrows, 1973), none of these phenotypes appears to be analogous to the Hms phenotype of Y . pestis.…”

Section: Discussionmentioning

confidence: 99%

“…Haemin-and CR-binding phenotypes serve distinct roles in a variety of pathogens (Daskaleros & Payne, 1987;Gardufio & Kay, 1992;Genco et al, 1994;Kay et al, 1985;Scott et al, 1993); the ability to bind these substrates has been correlated with virulence, but not always with haemin utilization. A long-held hypothesis is that the Hms' phenotype allows Y .…”

Section: Introductionmentioning

confidence: 99%

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The haemin storage (Hms+) phenotype of Yersinia pestis is not essential for the pathogenesis of bubonic plague in mammals

LillardJr

Bearden²,

Fetherston³

et al. 1999

Microbiology

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The haemin storage (Hms+) phenotype of Yersinia pestis enables this bacillus to form greenish/brown or red colonies on haemin or Congo Red agar plates, respectively, at 26 but not 37 O C . Escherichia coli strains that contain mutations in genes essential for siderophore biosynthesis, porphyrin generation and/or haemin transport remain unable to utilize exogenous haemin as a nutritional iron or porphyrin source when transformed with the cloned Y. pestis hmsHFRS locus. Further physiological analysis of the Hms+ phenotype of Y. pestis strain KIM6+ suggests that the haemin and inorganic iron stored by the Hms system was not used nutritionally under subsequent iron-deficient conditions. In vitro analysis of the bactericidal effects of hydrogen peroxide, superoxide and nitric oxide showed that Hms' Y. pestis cells, in certain cases, were more susceptible than the Hms+ parent cells to these reactive oxygen species a t 26 and/or 37 "C.In adherence assays, a higher percentage of Hms+ cells were associated with HeLa cells and normal human neutrophils, compared to Hms' cells. However, the Hms+ phenotype did not provide any additional protection against the killing effects of neutrophils. Finally, LD, , analysis in subcutaneously infected mice showed that an Hms' strain was slightly more virulent than Hms+, indicating that the Hms phenotype is not essential for the pathogenesis of bubonic plague in mammals.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The haemin storage (Hms+) phenotype of Yersinia pestis is not essential for the pathogenesis of bubonic plague in mammals

LillardJr

Bearden²,

Fetherston³

et al. 1999

Microbiology

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“…TO identify the components responsible for Congo red binding, whole cells of strain W50 grown in haemin-excess batch culture (at a concentration of 4 mg ml-l) were subjected to various pretreatments as described by Scott et al (1993). These included (a) papain in NaCl/Tris pH 6.5, containing 1.3 mM cysteine hydrochloride; (b) pepsin in 10 mM HC1; (c) trypsin in NaCl/Tris, pH 8.5 ; and (d) proteinase K in NaCl/Tris, pH 7-4.…”

Section: Congo Red Binding To Cells After Various Pretreatmentsmentioning

confidence: 99%

Congo red binding by Porphyromonas gingivalis is mediated by a 66 kDa outer-membrane protein

Smalley

Birss²,

McKee³

et al. 1995

Microbiology

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Congo red was bound from solution by strains of porlphyromonas gingivalis including W50, HG189, HG184, NCTC 11834, Bg 381, WPH35, the slower brown pigmenting colonial variant W50/BR1, and the avirulent mutant W50/BE1, and by Porphyromonas endodontalis HG370 and Porphyromonas asaccharolytica B537. SDS-PAGE of whole cells of all species examined displayed a 66 kDa Congo-red-binding component which was also detected in the outer membranes of P. gingivalis W50 grown in the chemostat under both haemin limitation and haemin excess, and which corresponded to a Coomassie-bluestained band of the same mobility. Pretreatment of haemin-excess batchgrown cells of P. gingiwalis W50 with polymyxin B, which binds to lipid A, did not inhibit binding, whilst binding was enhanced in the presence of 2 M ammonium sulphate, suggesting the involvement of non-specif ic hydrophobic interactions. Binding was also reduced by pretreatment with trypsin and papain, and by 8-anilino-l-naphthalenesulphonic acid, which binds to hydrophobic amino acids. The 66 kDa binding component was sensitive to proteinase K digestion, and loss of Congo red staining of this band correlated with the quantitative reduction in Congo red binding by whole cells. These data, and our previous work, show that Congo red and iron protoporphyrin IX (haemin) are bound to different outer-membrane components, and that Congo red binding may be of little value as a marker to detect virulent strains of P. gingiwalis or those expressing haemin-binding proteins.

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“…denticola does not appear to produce siderophores (45), so it must have other mechanisms for iron acquisition. Indeed, T. denticola can bind lactoferrin (50) and hemin (7,44).…”

mentioning

confidence: 99%

“…In addition, T. denticola produces a hemolysin (6), which could lyse erythrocytes in vivo and provide heme compounds for the bacterium. Previous studies have shown that T. denticola contains at least two hemin binding molecules: a 47-kDa constitutively produced hemin binding protein from strain ATCC 35405 (45) and a 44-kDa hemin binding protein (HbpA), which is up-regulated under iron-restricted growth, that is found in strains GM-1, MS-25, ATCC 33520, and ATCC 33404 (7). However, the role of these proteins in iron acquisition is unproven and the mechanism of binding and iron transport by these proteins has not been characterized.…”

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confidence: 99%

Cloning and Expression of Two Novel Hemin Binding Protein Genes fromTreponema denticola

Holt

Kolodrubetz

2001

Infect Immun

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Treponema denticola does not appear to produce siderophores, so it must acquire iron by other pathways. Indeed, T. denticola has been shown to have an iron-regulated 44-kDa outer membrane protein (HbpA) with hemin binding ability. To characterize the HbpA protein, its gene was cloned from genomic DNA libraries of T. denticola. Sequence analysis of the hbpA open reading frame indicated that it encoded a 42.8-kDa protein with a 23-amino-acid signal peptide. HbpA has no significant homology to any proteins in the databases. Southern blot analysis demonstrated that hbpA is present in several T. denticola ATCC strains and clinical isolates, but not in Treponema pectinovorum, Treponema socranskii, or Escherichia coli. HbpA, expressed as a recombinant protein in E. coli and purified by antibody affinity chromatography, has hemin binding activity as determined by lithium dodecyl sulfate-polyacrylamide gel electrophoresis with tetramethylbenzidine staining. Northern blot analysis showed that there were two hbpA-containing transcripts, of approximately 1.3 and 2.6 kb, and that the RNA levels were low-iron induced. Interestingly, the 2.6-kb mRNA also encoded a second protein with significant homology to hbpA. This downstream gene, called hbpB, was cloned and sequenced and its product was expressed as a fusion protein in E. coli. The hbpB gene product is 49% identical to HbpA and binds hemin. Thus, T. denticola has two novel hemin binding proteins which may be part of a previously unrecognized iron acquisition pathway.

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Binding of hemin and Congo red by oral hemolytic spirochetes

Cited by 22 publications

References 35 publications

The haemin storage (Hms+) phenotype of Yersinia pestis is not essential for the pathogenesis of bubonic plague in mammals

The haemin storage (Hms+) phenotype of Yersinia pestis is not essential for the pathogenesis of bubonic plague in mammals

Congo red binding by Porphyromonas gingivalis is mediated by a 66 kDa outer-membrane protein

Cloning and Expression of Two Novel Hemin Binding Protein Genes fromTreponema denticola

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