Iron-regulated outer membrane proteins in the periodontopathic bacterium, Bacteroides gingivalis

Bramanti, Thomas E.; Holt, Stanley C.

doi:10.1016/0006-291x(90)90986-w

Cited by 59 publications

(56 citation statements)

References 14 publications

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“…This pattern was similar to that described for the OM of strain W50 grown in haemin-restricted batch culture (Bramanti & Holt, 1990>, which was attributed by Kennel1 & Holt (1990) to the association of peptides with LPS cornponents. A high molecular mass component of R, z 0-02, which was resolved by densitometry as a doublet (Fig.…”

Section: Sds-page Polypeptide Projlessupporting

confidence: 88%

“…1). This finding was in keeping with the observations of Bramanti & Holt (1990), and necessitated solubilization of larger amounts of sample. Despite this, a faint ladder-like series of polypeptides was observed in the OM derived from haemin-limited cells (track 3), but not from those grown Fig.…”

Section: Sds-page Polypeptide Projlessupporting

confidence: 87%

“…Studies of P. gingiualis have shown that growth under haemin-limitation can lead to rapid changes in the profile of OMPs and to the induction of J . W. S m a k y and others novel proteins (Bramanti & Holt, 1990;Papaioannou et ul., 1991). The aim of the present study was to determine whether any of these OMPs were haemin-binding proteins.…”

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confidence: 99%

“…The aim of the present study was to determine whether any of these OMPs were haemin-binding proteins. In this study, in order to avoid the growth-raterelated changes in OMPs observed by Bramanti & Holt (1990) in batch culture, OMS were extracted from cells cultured in a chemostat at a constant growth rate and environmental pH, under either haemin-excess or haemin-limited conditions.…”

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confidence: 99%

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Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation

Smalley¹,

Birss²,

McKee³

et al. 1993

Journal of General Microbiology

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Porphyrornunns gingivalis W50 was grown in a chemostat at pH 7.3 under haemin-limitation and haemin-excess at a constant mean doubling time of 6-9 h. Outer membranes (OM) were extracted from whole cells using EDTA and compared by SDS-PAGE. Haemin-limited cells expressed novel outer membrane proteins (OMPs) of mol. mass 115, 113 and 19 kDa when samples were solubilized at 100 "C. A 46 kDa OMP was observed in haeminexcess cells but not in those from haemin-limited conditions. Tetramethylbenzidine (TMBZ) staining of gels, after OM solubilization at 20 "C, was used to detect haemin-binding proteins (HBPs). HBPs were observed only in OM from haemin-limited cells, The major HBP (mol, mass 32.4 kDa) corresponded to a similar sized Kenacid-bluestained protein which was not observed in haemin-excess-derived OM. Haemin-limited cells and OM displayed a ladder-like series of Kenacid-blue-stained proteins. Lighter TMBZ-stained proteins of mol. mass 51, 53, 56 and 60 kDa, with mobilities corresponding to those of silver-stained LPS components, were observed in haemin-limited OM. No soluble HBPs were detected extracellularly. The greater number of HBPs expressed by cells grown under haemin-limitation may reflect an additional cell surface receptor system for haemin acquisition under low environmental levels of this essential cofactor.

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Section: Sds-page Polypeptide Projlessupporting

confidence: 88%

Section: Sds-page Polypeptide Projlessupporting

confidence: 87%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation

Smalley¹,

Birss²,

McKee³

et al. 1993

Journal of General Microbiology

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“…Whole cells of P. gingivalis can bind haemin (Smalley e t al., 1991(Smalley e t al., , 1993(Smalley e t al., , 1994Carman e t al., 1990;Genco et al, 1994) via the lipid A fraction of lipopolysaccharide (Grenier, 1991). In addition, novel outer-membrane proteins are expressed under haemin limitation (Bramanti & Holt, 1990, 1992aPapaioannou et al, 1991), some of which are haeminbinding proteins (Bramanti & Holt, 1993;Smalley e t al., 1993). Although the growth of members of the Porph_yromonas group in vitro is inhibited to varying degrees by Congo red (Mayrand et al, 1984) it is not known whether P. gingivalis binds this dye.…”

Section: Introductionmentioning

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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Detection and identification of bacterial cell surface proteins by fluorescent labeling

2007

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A rapid method of detection and identification of bacterial cell surface proteins is needed to better understand the interaction of bacteria with host components. To detect cell surface proteins, we have labeled cells of the Gram-negative anaerobic bacterium, Porphyromonas gingivalis, with fluorescent cyanine dyes, Cy3 and Cy5. We demonstrate that only cell surface proteins were labeled, indicating the method applied in our study is suitable for detection and identification of cell surface proteins in Gram-negative bacteria and possibly other organisms.

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Iron-regulated outer membrane proteins in the periodontopathic bacterium, Bacteroides gingivalis

Cited by 59 publications

References 14 publications

Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation

Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation

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

Detection and identification of bacterial cell surface proteins by fluorescent labeling

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