ARDC Research Link Australia

Publication

Publisher: Elsevier BV

Date: 12-2020

DOI: 10.1016/J.DENTAL.2020.09.022

Publication

Complete Nucleotide Sequence of a Gene prtR of Porphyromonas gingivalis W50 Encoding a 132-kDa Protein That Contains an Arginine-Specific Thiol Endopeptidase Domain and a Hemagglutinin Domain

Publisher: Elsevier BV

Date: 02-1995

DOI: 10.1006/BBRC.1995.1205

Abstract: We have purified from Porphyromonas gingivalis W50 a 45 kDa arginine-specific, thiol-activated, EDTA-sensitive endopeptidase, designated prtR. Oligonucleotide probes based on the N-terminal amino acid sequence were used to isolate a genomic fragment containing an open reading frame (3654 bp) with the potential to encode a 132 kDa protein including the prtR N-terminus. Analysis of this prtR gene revealed that the predicted nascent product contains a protease domain followed by a haemagglutinin domain and is post-translationally processed by proteolytic (possibly autolytic) events to produce a 43-54 kDa arginine-specific, thiol protease and a 41-53 kDa haemagglutinin. Comparison of the prtR with the P. gingivalis prtH gene suggests that the prtH gene product also contains protease and haemagglutinin domains but in the reverse order to that in the prtR. An overlapping but shifted reading frame at the 3' end of the prtR encodes the 5' region of the prtH.

Publication

An ELISA for a multiphosphorylated peptide, α(s1)-casein(59-79)

Publisher: Elsevier BV

Date: 05-1998

DOI: 10.1016/S0022-1759(98)00043-X

Abstract: Multiphosphorylated segments of proteins are predicted to be in or near epitopes. However, due to the very hydrophilic nature of multiphosphorylated peptides, epitope mapping by ELISA using conventional microtitre plates can produce false negatives due to poor antigen adsorption. We have developed a sensitive ELISA for a multiphosphorylated peptide alpha(s1)-casein(59-79) containing five phosphoseryl residues using Nunc-Immuno Maxisorp modules for antigen adsorption. The peptide alpha(s1)-casein(59-79) was detected in the ELISA at antigen coating concentrations of 1.0 microg/ml and above, using rabbit anti-casein antibodies at a dilution of 1/10,000 in Tris-buffered saline containing 0.05% (w/v) Tween 20 and 1.0% (v/v) normal goat serum. At an antigen coating concentration of 10 microg/ml, anti-casein antibodies bound to alpha(s1)-casein(59-79) and produced an absorbance of more than 100 times background. Using conventional polyvinyl chloride and polystyrene plates the peptide alpha(s1)-casein(59-79) could only be detected at very high antigen coating concentrations of 1-10 mg/ml. The addition of 0.05% (w/v) Tween 20 to the blocking, antibody diluting and wash buffers of the ELISA was shown to significantly reduce nonspecific binding of the primary antibody. Further, the inclusion of normal goat serum in the blocking and antibody diluting buffers resulted in a small but significant increase in absorbance. The ELISA developed in this study has been used successfully with a range of enzymatically derived and synthetic peptides containing one to five phosphorylated residues such that it should have general applicability to the study of antigenicity of multiphosphorylated peptides.

Publication

Publisher: Wiley

Date: 12-11-2021

DOI: 10.1111/JCPE.13389

Publication

The synthesis of phosphopeptides via the Bpoc-based approach

Publisher: Royal Society of Chemistry (RSC)

Date: 2007

DOI: 10.1039/B617699M

Abstract: The 2-(p-biphenylyl)-2-propyloxycarbonyl (Bpoc) group was examined as an N(alpha)-protecting group in the stepwise assembly of the MAP Kinase ERK2 [178-188 Thr(P)(183), Tyr(P)(185)] peptide. The mild acid deprotection of the Bpoc group permitted (i) incorporation of a fully protected phosphothreonyl derivative and (ii) a TFA-based final cleavage step. The first five C-terminal residues (184-188) were incorporated in the Fmoc mode of peptide synthesis, with all subsequent amino acids coupled as their Bpoc-Xxx-OH derivatives. The target product was obtained in high purity and yield, indicating that a Bpoc-based approach to phosphopeptide synthesis was compatible with both the acid-labile side chain protecting groups employed and Hmp-Wang resin.

Publication

Effect of ozone and Tooth Mousse TM on the efficacy of peroxide bleaching

Publisher: Wiley

Date: 21-05-2008

DOI: 10.1111/J.1834-7819.2008.00021.X

Abstract: The aim of this in vitro study was to investigate the effect of Tooth Mousse and ozone on the bleaching effectiveness of peroxide (P). Sixty enamel specimens were stained by tea infusion. P (8% carbamide peroxide solution) and the P/TM (50:50) blend were prepared freshly as required. The specimens were ided randomly into six groups: Group A - ozone followed by P Group B - ozone concurrently with P Group C - P alone Group D - ozone followed by P/TM Group E - ozone concurrently with P/TM and Group F - P/TM alone. Ozone exposure was of 40 seconds duration. Digital photographic images were recorded at baseline and endpoint under standardized lighting and desiccation conditions. CIELAB L*a*b* values were determined. The addition of TM to P or the application of ozone with P did not significantly affect bleaching effectiveness compared with P alone. The application of ozone prior to P significantly decreased bleaching effectiveness as indicated by the DeltaL*, Deltaa*, DeltaE and %L* values. The addition of TM to the P did enhance the aesthetic by increasing the lustre and translucency of the treated enamel. The results of this study suggest that Tooth Mousse may be applied concurrently with the bleach, and not reduce bleaching effectiveness.

Publication

Ion release from calcium and fluoride containing dental varnishes.

Publisher: Wiley

Date: 04-02-2014

DOI: 10.1111/ADJ.12144

Abstract: A range of dental varnishes have been commercialized recently that contain calcium and inorganic phosphate in addition to fluoride. The aim of this study was to analyse the fluoride, calcium and inorganic phosphate ion release from: (1) MI Varnish containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (2) Clinpro White containing functionalized tricalcium phosphate (fTCP) (3) Enamel Pro containing amorphous calcium phosphate (4) Bifluorid 5 containing calcium fluoride and (5) Duraphat (no added calcium control). The varnishes were applied to a standardized surface area of polyvinyl chloride (n = 7 per group) and immersed in 25 g of distilled deionized water which was changed at 1, 4, 24, 72 and 168 hours. The ion release was determined by ion chromatography and expressed as μmol (cumulative) per gram of varnish. All varnishes released measurable fluoride and calcium, however only MI Varnish and Enamel Pro released significant levels of inorganic phosphate. At 24 hours the order of cumulative fluoride release was: 1>3>4>2=5 with 1 significantly higher (p 4>3>2=5 with 1 significantly higher (p < 0.05) than the rest. MI Varnish containing CPP-ACP had the highest release of calcium and fluoride ions.

Publication

Characterization of the O-Glycoproteome of Tannerella forsythia

Publisher: American Society for Microbiology

Date: 27-10-2021

DOI: 10.1128/MSPHERE.00649-21

Abstract: Tannerella forsythia is an oral pathogen associated with severe forms of periodontal disease characterized by destruction of the tooth’s supporting tissues, including the bone. The bacterium releases a variety of proteins associated with virulence on the surface of outer membrane vesicles.

Publication

Publisher: Elsevier BV

Date: 11-2023

DOI: 10.1016/J.MOLMED.2023.08.006

Publication

Interplay between Porphyromonas gingivalis and EGF signalling in the regulation of CXCL14.

Publisher: Hindawi Limited

Date: 23-03-2018

DOI: 10.1111/CMI.12837

Abstract: Porphyromonas gingivalis is a keystone pathogen in chronic periodontitis. Its expression of gingipain proteases (Kgp and RgpA/B) is central to the stimulation of chronic inflammation. In this study, we investigated the inflammatory response of oral epithelial cells to P. gingivalis. The cells responded by upregulating the expression of the orphan chemokine CXCL14. The stimulation of CXCL14 expression was largely triggered by the gingipain proteases and was dependent on the host protease-activated receptor PAR-3. Significantly, CXCL14 expression was transcriptionally repressed in response to epidermal growth factor (EGF)-induced activation of the MEK-ERK1/2 pathway. P. gingivalis overcomes the repression of CXCL14 via the gingipain protease-mediated degradation of EGF. Therefore, P. gingivalis not only directly stimulates CXCL14 expression via PAR-3 but also promotes its expression by antagonising EGF signalling. In addition to chemotactic activity, some chemokines also have antimicrobial activities. CXCL14 was demonstrated to have bactericidal activity, against commensal oral streptococci associated with health. Notably though, P. gingivalis was not susceptible to killing by CXCL14, potentially because the gingipain proteases can degrade CXCL14. This suggests that the stimulation of dysregulated CXCL14 expression by P. gingivalis may help promote dysbiosis and the development of chronic periodontitis.

Publication

Resin infiltration of developmentally hypomineralised enamel

Publisher: Wiley

Date: 15-02-2013

DOI: 10.1111/IPD.12025

Abstract: Molar incisor hypomineralisation (MIH) is a problematic condition with several characteristics for which infiltrant resins could theoretically improve clinical outcomes. To investigate whether caries infiltrant resin can penetrate MIH-affected enamel. Molar incisor hypomineralisation lesions (n = 21) were infiltrated using either the standard protocol or with the addition of a sodium hypochlorite (NaOCl) irrigation step. Lesions were sectioned and examined microscopically for infiltrant penetration before undergoing Vickers hardness testing. The surfaces of several lesions were also examined using scanning electron microscopy (SEM). Infiltrant resin could penetrate MIH lesions however, the pattern was erratic. Two lesions were confined to inner enamel, and no infiltration occurred. On average, the resin penetrated to a depth of 0.67 ± 0.39 mm and 23.1 ± 15.2% of the area of the lesion. Microhardness increased in areas of resin penetration by 1.0 ± 0.7 GPa representing a proportional increase of 2.2 ± 2.5 times. There were no significant differences in results based on either the infiltration protocol or the type of MIH lesion. Caries infiltrant resin is capable of penetrating MIH enamel lesions however, the pattern, extent, and change in hardness produced are currently unpredictable.

Publication

Streptococcus mutans biofilm disruption by κ-casein glycopeptide.

Publisher: Elsevier BV

Date: 06-2013

DOI: 10.1016/J.JDENT.2013.03.010

Abstract: Caseinomacropeptide (CMP), the variably phosphorylated and glycosylated forms of the bovine milk protein fragment, κ-casein(106-169), is produced during cheese production and has been shown to have a range of antibacterial bioactivities. To characterise the biofilm disruptive component of CMP and compare its activity with the known antimicrobial agents chlorhexidine and zinc ions. Streptococcus mutans biofilms were grown in flow cells with an artificial saliva medium containing sucrose and treated with CMP and the glycosylated forms of κ-casein(106-169) (κ-casein glycopeptide, KCG). The biofilms were imaged using confocal laser scanning microscopy (CLSM) and quantified by COMSTAT software analysis. A static biofilm assay and flow cytometric analysis were used to examine the mechanism of action of chlorhexidine and a combination of KCG with the known antimicrobial agent ZnCl2 (KCG-Zn). CLSM analysis showed that S. mutans produced robust, structured biofilms with an average thickness of 7.37μm and a biovolume of 3.88μm(3)/μm(2) substratum after 16h of incubation in the flow cell system. A single application of 10mg/mL CMP that contained 2.4mg/mL KCG significantly reduced total biofilm biovolume and average biofilm thickness by 53% and 61%, respectively. This was statistically the same as a 2.4mg/mL KCG treatment that reduced the total biovolume and average thickness by 59% and 69%, respectively, suggesting the KCG was the biofilm disruptive component of CMP. Chlorhexidine treatment (0.1%) caused similar effects in the flow cell model. KCG-Zn caused significantly more disruption of the biofilms than either KCG or ZnCl2 treatment alone. In a static biofilm model chlorhexidine was shown to work by disrupting bacterial membrane integrity whilst KCG-Zn had no effect on membrane integrity. KCG and KCG-Zn may have potential as natural biofilm disruptive agents.

Publication

Combined proteomic and transcriptomic interrogation of the venom gland of Conus geographus uncovers novel components and functional compartmentalization.

Publisher: Elsevier BV

Date: 04-2014

DOI: 10.1074/MCP.M113.031351

Publication

PG0026 Is the C-terminal Signal Peptidase of a Novel Secretion System of Porphyromonas gingivalis

Publisher: Elsevier BV

Date: 07-2012

DOI: 10.1074/JBC.M112.369223

Publication

Publisher: Elsevier BV

Date: 07-2014

DOI: 10.1074/JBC.M114.584540

Publication

Publisher: Wiley

Date: 23-04-2004

DOI: 10.1111/J.0906-6713.2004.003559.X

Publication

Sodium ion-driven serine/threonine transport in Porphyromonas gingivalis.

Publisher: American Society for Microbiology

Date: 15-07-2001

DOI: 10.1128/JB.183.14.4142-4148.2001

Abstract: Porphyromonas gingivalis is an asaccharolytic, gram-negative bacterium that relies on the fermentation of amino acids for metabolic energy. When grown in continuous culture in complex medium containing 4 mM (each) free serine, threonine, and arginine, P. gingivalis assimilated mainly glutamate/glutamine, serine, threonine, aspartate/asparagine, and leucine in free and/or peptide form. Serine and threonine were assimilated in approximately equal amounts in free and peptide form. We characterized serine transport in this bacterium by measuring uptake of the radiolabeled amino acid in washed cells of P. gingivalis energized with a tetrapeptide not containing serine. Serine was transported by a single system with an affinity constant for transport ( K t ) of 24 μM that was competitively inhibited by threonine. Serine transport was dependent on sodium ion concentration in the suspending buffer, and the addition of the ionophore gramicidin caused the inhibition of serine uptake. Together these data indicate that serine transport was sodium ion-motive force driven. A P. gingivalis gene potentially encoding a serine transporter was identified by sequence similarity to an Escherichia coli serine transporter (SstT). This P. gingivalis gene, designated sstT, was inactivated by insertion of a Bacteroides tetQ gene, producing the mutant W50ST. The mutant was unable to transport serine, confirming the presence of a single serine transporter in this bacterium under these growth conditions. The transport of serine by P. gingivalis was dependent on the presence of free cysteine in the suspension buffer. Other reducing agents were unable to stimulate serine uptake. These data show that P. gingivalis assimilates free serine and threonine from culture media via a cysteine-activated, sodium ion-motive force-driven serine/threonine transporter.

Publication

Publisher: Elsevier

Date: 2013

DOI: 10.1016/B978-0-12-382219-2.00308-2

Publication

Publisher: Springer Science and Business Media LLC

Date: 25-04-2007

DOI: 10.1007/S10989-007-9094-Z

Publication

Identification of a novel heterodimeric outer membrane protein of Porphyromonas gingivalis by two‐dimensional gel electrophoresis and peptide mass fingerprinting

Publisher: Wiley

Date: 09-2001

DOI: 10.1046/J.1432-1327.2001.02399.X

Abstract: Porphyromonas gingivalis is a Gram-negative, anaerobic bacterium associated with chronic periodontitis. A 2D electrophoretic analysis of the outer membrane of P. gingivalis W50 revealed a dominant train of spots at 40-41 kDa. The proteins in the train of spots were digested in-gel with trypsin and identified by MS. The train of spots represented two proteins, designated Omp40 and Omp41 that share 47% sequence identity. Preparation of outer membranes in the absence of protease inhibitors resulted in partial cleavage of Omp40 and Omp41 to produce an N-terminal and C-terminal fragment of both proteins. The N-terminal fragments displayed the same isoelectric heterogeneity as the intact proteins. Almost 100% of the amino-acid sequence of these N-terminal fragments in each 2D gel spot was verified suggesting lack of post-translational modification. Re-subjecting a single N-terminal domain spot to 2D electrophoresis resulted in the complete series of spots being reproduced, suggesting that the heterogeneity was related to conformational equilibria. Under reduced conditions and without heating, Omp40 and Omp41 migrated as 34- to 35-kDa proteins in SDS/PAGE whereas under nonreduced conditions the proteins migrated as 70-kDa proteins, suggesting the formation of dimers through intersubunit disulfide bonds. The proteins each contain two cysteine residues in the conserved sequence RPVSCPECPE. Tryptic peptides generated from the nonreduced forms of the proteins confirmed the presence of heterodimers stabilized through intersubunit disulfide bond formation. With the exception of heterodimer formation, the two proteins share several similarities with OmpA-like porins of other Gram-negative bacteria including consensus sequence, abundance, modification by heat, overall length and positioning of domains.

Publication

Virulence factors of the oral spirochete Treponema denticola

Publisher: SAGE Publications

Date: 12-10-2011

DOI: 10.1177/0022034510385242

Abstract: There is compelling evidence that treponemes are involved in the etiology of several chronic diseases, including chronic periodontitis as well as other forms of periodontal disease. There are interesting parallels with other chronic diseases caused by treponemes that may indicate similar virulence characteristics. Chronic periodontitis is a polymicrobial disease, and recent animal studies indicate that co-infection of Treponema denticola with other periodontal pathogens can enhance alveolar bone resorption. The bacterium has a suite of molecular determinants that could enable it to cause tissue damage and subvert the host immune response. In addition to this, it has several non-classic virulence determinants that enable it to interact with other pathogenic bacteria and the host in ways that are likely to promote disease progression. Recent advances, especially in molecular-based methodologies, have greatly improved our knowledge of this bacterium and its role in disease.

Publication

Role of RgpA, RgpB, and Kgp proteinases in virulence of Porphyromonas gingivalis W50 in a murine lesion model.

Publisher: American Society for Microbiology

Date: 12-2001

DOI: 10.1128/IAI.69.12.7527-7534.2001

Abstract: Extracellular Arg-x- and Lys-x-specific cysteine proteinases are considered important virulence factors and pathogenic markers for Porphyromonas gingivalis , a bacterium implicated as a major etiological agent of chronic periodontitis. Three genes. rgpA , rgpB, and kgp, encode an Arg-x-specific proteinase and adhesins (RgpA), an Arg-x-specific proteinase (RgpB), and a Lys-x-specific proteinase and adhesins (Kgp), respectively. The contribution to pathogenicity of each of the proteinase genes of P. gingivalis W50 was investigated in a murine lesion model using isogenic mutants lacking RgpA, RgpB, and Kgp. Whole-cell Arg-x-specific proteolytic activity of both the RgpA − and RgpB − isogenic mutants was significantly reduced (3- to 4-fold) relative to that of the wild-type W50. However, for the Kgp − isogenic mutant, whole-cell Arg-x activity was similar to that of the wild-type strain. Whole-cell Lys-x proteolytic activity of the RgpA − and RgpB − mutants was not significantly different from that of wild-type W50, whereas the Kgp − mutant was devoid of Lys-x whole-cell proteolytic activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis using proteinase-specific antibodies of cell sonicates of the wild-type and mutant strains showed that the proteinase catalytic domain of each of the mutants was not expressed. This analysis further showed that RgpB appeared as 72- and 80-kDa bands, and the catalytic domains of RgpA and Kgp appeared as processed 45-kDa and 48-kDa bands, respectively. In the murine lesion model, mice were challenged with three doses of each mutant and wild-type strain. At the lower dose (3.0 × 10 9 viable-cells), no lesions were recorded for each of the mutants, whereas wild-type W50 induced large ulcerative lesions. At a dose of 6.0 × 10 9 viable-cells, all the mice challenged with the wild-type strain died, whereas mice challenged with the RgpA − and RgpB − isogenic mutants did not die but developed lesions. Mice challenged with the Kgp − isogenic mutant at this dose did not develop lesions. At a 1.2 × 10 10 viable-cell dose, only 40% of mice challenged with the Kgp − mutant developed lesions, and these lesions were significantly smaller than lesions induced by the wild-type strain at the 3.0 × 10 9 viable-cell dose. All the mice challenged with the RgpA − mutant died at the 1.2 × 10 10 viable-cell dose, whereas only 20% died when challenged with the RgpB − mutant at this dose. Wild-type phenotype was restored to the RgpB − mutant by complementation with plasmid pNJR12:: rgpB containing the rgpB gene. There was no difference between the pNJR12:: rgpB -complemented RgpB − mutant and the wild-type W50 strain in whole-cell Arg-x activity, protein profile, or virulence in the murine lesion model. These results show that the three proteinases, RgpA, RgpB, and Kgp, all contributed to virulence of P. gingivalis W50 in the murine lesion model and that the order in which they contributed was Kgp ≫ RgpB ≥ RgpA.

Publication

Fluoride and casein phosphopeptide-amorphous calcium phosphate.

Publisher: SAGE Publications

Date: 04-2008

DOI: 10.1177/154405910808700420

Abstract: Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) slows the progression of caries and remineralizes enamel subsurface lesions. The aim of this study was to determine the ability of CPP-ACP to increase the incorporation of fluoride into plaque and to promote enamel remineralization in situ. Randomized, double-blind, cross-over studies involved mouthrinses and dentifrices containing CPP-ACP and fluoride. The mouthrinses were used for 60 sec, three times/day for 5 days, and supragingival plaque was collected and analyzed for F. The dentifrices were rinsed as a water slurry for 60 sec four times/day for 14 days in an in situ model. The addition of 2% CPP-ACP to the 450-ppm-F mouthrinse significantly increased the incorporation of fluoride into plaque. The dentifrice containing 2% CPP-ACP produced a level of remineralization similar to that achieved with a dentifrice containing 2800 ppm F. The dentifrice containing 2% CPP-ACP plus 1100 ppm F was superior to all other formulations.

Publication

Publisher: Public Library of Science (PLoS)

Date: 26-08-2013

DOI: 10.1371/JOURNAL.PONE.0071727

Publication

Type IX Secretion System Cargo Proteins Are Glycosylated at the C Terminus with a Novel Linking Sugar of the Wbp/Vim Pathway

Publisher: American Society for Microbiology

Date: 27-10-2020

DOI: 10.1128/MBIO.01497-20

Abstract: Porphyromonas gingivalis and Tannerella forsythia , two pathogens associated with severe gum disease, use the type IX secretion system (T9SS) to secrete and attach toxic arrays of virulence factor proteins to their cell surfaces. The proteins are tethered to the outer membrane via glycolipid anchors that have remained unidentified for more than 2 decades. In this study, the first sugar molecules (linking sugars) in these anchors are identified and found to be novel compounds. The novel biosynthetic pathway of these linking sugars is also elucidated. A erse range of bacteria that do not have the T9SS were found to have the genes for this pathway, suggesting that they may synthesize similar linking sugars for utilization in different systems. Since the cell surface attachment of virulence factors is essential for virulence, these findings reveal new targets for the development of novel therapies.

Publication

Remineralization of enamel subsurface lesions by chewing gum with added calcium

Publisher: Elsevier BV

Date: 10-2009

DOI: 10.1016/J.JDENT.2009.06.003

Abstract: Chewing sugar-free gum has been shown to promote enamel remineralization. Manufacturers are now adding calcium to the gum in an approach to further promote enamel remineralization. The aim of this study was to compare the remineralization efficacy of four sugar-free chewing gums, two containing added calcium, utilizing a double-blind, randomized, crossover in situ model. The sugar-free gums were: Trident Xtra Care, Orbit Professional, Orbit and Extra. Ten subjects wore removable palatal appliances with four human-enamel half-slab insets containing subsurface demineralized lesions. For four times a day for 14 consecutive days subjects chewed one of the chewing gums for 20min. After each treatment the enamel slabs were removed, paired with their respective demineralized control slabs, embedded, sectioned and mineral level determined by microradiography. After 1-week rest the subjects chewed another of the four gums and this was repeated until each subject had used the four gum products. Chewing with Trident Xtra Care resulted in significantly higher remineralization (20.67+/-1.05%) than chewing with Orbit Professional (12.43+/-0.64%), Orbit (9.27+/-0.59%) or Extra (9.32+/-0.35%). The form of added calcium in Trident Xtra Care was CPP-ACP and that in Orbit Professional calcium carbonate with added citric acid/citrate for increased calcium solubility. Although saliva analysis confirmed release of the citrate and calcium from the Orbit Professional gum the released calcium did not result in increased enamel remineralization over the normal sugar-free gums. These results highlight the importance of calcium ion bioavailability in the remineralization of enamel subsurface lesions in situ.

Publication

Bioinformatic investigation of the cost management strategies of five oral microbes.

Publisher: Wiley

Date: 09-10-2015

DOI: 10.1111/OMI.12071

Abstract: Some amino acids are more energetically costly to synthesize de novo, therefore many microbes have evolved to regulate the metabolic expenditure of the cell and reduce the energy burden of extracellular unrecyclable proteins. Several oral bacterial species take up amino acids and peptides obtained from proteolysis of host proteins and hence do not rely only on de novo synthesis. The aim of this study was to investigate if five oral bacterial species implement cost management strategies to reduce the energy burden of extracellular unrecyclable proteins. Since the relative de novo amino acid synthesis costs are proportional to the masses of the amino acids, the energy costs of producing proteins were assessed by calculating the mean amino acid mass for each protein. For Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Prevotella intermedia and Streptococcus sanguinis, the outer membrane/extracellular proteins are made up of a much larger percentage of lower average mass amino acids whereas cytoplasmic proteins are made up of a larger proportion of higher average mass amino acid residues. These results are consistent with the five oral bacterial species employing energy-saving mechanisms in the production of extracellular unrecyclable proteins. Interestingly, the P. gingivalis and S. sanguinis genomes exhibited significantly lower predicted mean amino acid masses compared with those of the genomes of the other three species, suggesting that this may provide them with an energy advantage with respect to protein biosynthetic cost.

Publication

C‐Terminal Modification and Multimerization Increase the Efficacy of a Proline‐Rich Antimicrobial Peptide

Publisher: Wiley

Date: 16-11-2016

DOI: 10.1002/CHEM.201604172

Abstract: Two series of branched tetramers of the proline-rich antimicrobial peptide (PrAMP), Chex1-Arg20, were prepared to improve antibacterial selectivity and potency against a panel of Gram-negative nosocomial pathogens including Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa. First, tetramerization was achieved by dithiomaleimide (DTM) conjugation of two C-terminal-cysteine bearing dimers that also incorporated C-terminal peptide chemical modification. DTM-linked tetrameric peptides containing a C-terminal hydrazide moiety on each dimer exhibited highly potent activities in the minimum inhibitory concentration (MIC) range of 0.49-2.33 μm. A second series of tetrameric analogues with C-terminal hydrazide modification was prepared by using alternative conjugation linkers including trans-1,4-dibromo-2-butene, α,α'-dibromo-p-xylene, or 6-bismaleimidohexane to determine the effect of length on activity. Each displayed potent and broadened activity against Gram-negative nosocomial pathogens, particularly the butene-linked tetrameric hydrazide. Remarkably, the greatest MIC activity is against P. aeruginosa (0.77 μm/8 μg mL

Publication

Errata

Publisher: Wiley

Date: 26-05-2014

DOI: 10.1111/ADJ.12185

Publication

Physicochemical Characterization of Casein Phosphopeptide-Amorphous Calcium Phosphate Nanocomplexes

Publisher: Elsevier BV

Date: 04-2005

DOI: 10.1074/JBC.M413504200

Publication

Publisher: Elsevier BV

Date: 09-1992

DOI: 10.1016/S0960-894X(00)80637-X

Publication

Publisher: Elsevier BV

Date: 05-2012

DOI: 10.1016/J.IDAIRYJ.2011.12.009

Publication

Importance of bioavailable calcium in fluoride dentifrices for enamel remineralization.

Publisher: Elsevier BV

Date: 11-2018

DOI: 10.1016/J.JDENT.2018.08.005

Abstract: To compare remineralization of enamel subsurface lesions by fluoride dentifrices with added calcium in a double-blind, randomized, cross-over, in situ study. Human enamel with subsurface lesions were prepared and inserted into intra-oral appliances worn by volunteers. A slurry (1 g toothpaste/4 ml H All calcium and fluoride containing toothpastes released > 90% of bioavailable fluoride and were superior to the respective fluoride alone toothpastes in remineralization of enamel subsurface lesions. The level of remineralization followed the order: CPP-ACP/1l00 ppm F > ACP/1150 ppm F = TCP/5000 ppm F > 5000 ppm F = CaCO Bioavailable calcium in fluoride dentifrices enhanced remineralization of enamel subsurface lesions.

Publication

Increased remineralization of tooth enamel by milk containing added casein phosphopeptide-amorphous calcium phosphate.

Publisher: Cambridge University Press (CUP)

Date: 24-01-2006

DOI: 10.1017/S0022029905001482

Abstract: Casein phosphopeptide amorphous calcium phosphate nanocomplexes (CPP-ACP) in chewing gum, lozenges and mouthrinses have been shown to remineralize enamel subsurface lesions in human in situ experiments. The aim of this double-blind, randomized clinical study was to investigate the capacity of CPP-ACP added to bovine milk to remineralize enamel subsurface lesions in situ . Ten subjects drank milk containing either 2·0 or 5·0 g CPP-ACP/l or a control milk whilst wearing removable appliances with enamel slabs containing subsurface demineralized lesions. Each 200 ml milk s le was consumed once a day for each weekday over three consecutive weeks. After each treatment and one weeks rest the subjects crossed over to the other treatments. At the completion of the treatments the enamel slabs were removed and remineralization determined using microradiography and microdensitometry. The results demonstrated that all three milk s les remineralized enamel subsurface lesions. However, the milk s les containing CPP-ACP produced significantly greater remineralization than the control milk. The remineralising effect of CPP-ACP in milk was dose-dependent with 2·0 and 5·0 g CPP-ACP/l producing an increase in mineral content of 70 and 148%, respectively, relative to the control milk. The differences in remineralization following exposure to the three milk s les were all statistically significant ( P ·001). In conclusion, this study shows that the addition of 2·0–5·0 g CPP-ACP/l to milk substantially increases its ability to remineralize enamel subsurface lesions.

Publication

Publisher: Elsevier BV

Date: 06-2002

DOI: 10.1074/JBC.M200811200

Publication

Bacterial membrane vesicles transport their DNA cargo into host cells

Publisher: Springer Science and Business Media LLC

Date: 08-2017

DOI: 10.1038/S41598-017-07288-4

Abstract: Bacterial outer membrane vesicles (OMVs) are extracellular sacs containing biologically active products, such as proteins, cell wall components and toxins. OMVs are reported to contain DNA, however, little is known about the nature of this DNA, nor whether it can be transported into host cells. Our work demonstrates that chromosomal DNA is packaged into OMVs shed by bacteria during exponential phase. Most of this DNA was present on the external surfaces of OMVs, with smaller amounts located internally. The DNA within the internal compartments of Pseudomonas aeruginosa OMVs were consistently enriched in specific regions of the bacterial chromosome, encoding proteins involved in virulence, stress response, antibiotic resistance and metabolism. Furthermore, we demonstrated that OMVs carry DNA into eukaryotic cells, and this DNA was detectable by PCR in the nuclear fraction of cells. These findings suggest a role for OMV-associated DNA in bacterial-host cell interactions and have implications for OMV-based vaccines.

Publication

Mass spectrometric analyses of peptides and proteins in human gingival crevicular fluid

Publisher: American Chemical Society (ACS)

Date: 22-01-2010

DOI: 10.1021/PR900775S

Abstract: Gingival crevicular fluid (GCF) is a pathophysiological fluid that flows into the oral cavity. Human GCF was collected using sterile glass microcapillary tubes from inflamed periodontal sites in patients who had a history of periodontal disease and were in the maintenance phase of treatment. S les from in idual sites were analyzed using MS techniques both before and following HPLC. GCF s les were also pooled and subjected to SDS-PAGE, in-gel digestion and MS analyses using both MALDI-TOF/TOF MS and nanoLC-ESI-MS/MS. MS spectra were used to search human protein sequence databases for protein identification. With these approaches, 33 peptides and 66 proteins were positively identified in human GCF. All of the peptides discovered in this study are reported in GCF here for the first time. Forty-three of the identified proteins, such as actin and the actin binding proteins profilin, cofilin and gelsolin, have not been reported in GCF before.

Publication

The Role of Multiphosphorylated Peptides in Mineralized Tissue Regeneration

Publisher: Springer Science and Business Media LLC

Date: 18-08-2007

DOI: 10.1007/S10989-007-9105-0

Publication

Protein substrates of a novel secretion system are numerous in the Bacteroidetes phylum and have in common a cleavable C-terminal secretion signal, extensive post-translational modification, and cell-surface attachment

Publisher: American Chemical Society (ACS)

Date: 23-09-2013

DOI: 10.1021/PR400487B

Abstract: The secretion of certain proteins in Porphyromonas gingivalis is dependent on a C-terminal domain (CTD). After secretion, the CTD is cleaved prior to extensive modification of the mature protein, probably with lipopolysaccharide, therefore enabling attachment to the cell surface. In this study, bioinformatic analyses of the CTD demonstrated the presence of three conserved sequence motifs. These motifs were used to construct Hidden Markov Models (HMMs) that predicted 663 CTD-containing proteins in 21 fully sequenced species of the Bacteroidetes phylum, while no CTD-containing proteins were predicted in species outside this phylum. Further HMM searching of Cytophaga hutchinsonii led to a total of 171 predicted CTD proteins in that organism alone. Proteomic analyses of membrane fractions and culture fluid derived from P. gingivalis and four other species containing predicted CTDs (Parabacteroides distasonis, Prevotella intermedia, Tannerella forsythia, and C. hutchinsonii) demonstrated that membrane localization, extensive post-translational modification, and CTD-cleavage were conserved features of the secretion system. The CTD cleavage site of 10 different proteins from 3 different species was determined and found to be similar to the cleavage site previously determined in P. gingivalis, suggesting that homologues of the C-terminal signal peptidase (PG0026) are responsible for the cleavage in these species.

Publication

Publisher: Wiley

Date: 05-1999

DOI: 10.1002/(SICI)1099-1387(199905)5:5<221::AID-PSC182>3.0.CO;2-O

Publication

Characterisation of developmentally hypomineralised human enamel

Publisher: Elsevier BV

Date: 07-2013

DOI: 10.1016/J.JDENT.2013.05.002

Abstract: To investigate and clarify physical and chemical properties of enamel affected by molar incisor hypomineralisation (MIH). A series of in vitro studies were performed on extracted molars affected by MIH and sound teeth for controls. Tooth sections underwent Vickers microhardness testing before lapping and subsequent transverse microradiographic analysis and examination under polarised light microscopy. Carbonate content was determined by CO2 release from acid digestion. Unprepared and fractured surfaces were examined under scanning electron microscopy. MIH-affected molars demonstrated a severe degree of hypomineralisation with an average mineral content of only 58.8%vol% mineral. Vickers microhardness was significantly reduced in MIH compared with controls (1.8±1.1 v 4.4±1.0 GPa, p<0.05) and polarised light microscopy revealed the bulk of MIH lesions had a porosity of ≤5% but also substantial areas of ≥10% and smaller areas exceeding 25% porosity. A surface layer was frequently observed on both intact and broken-down lesions and cervical regions of MIH teeth were typically spared. Carbonate content of MIH enamel was higher than control s les (6.6±2.1 v 4.4±1.1 wt%, p<0.05). Scanning electron microscopy showed that both the enamel rod and surface ultrastructure were defective. Clinical characteristics did not consistently correlate with all properties. The properties of MIH-affected enamel significantly differ from those of normal enamel and were highly variable, however some common characteristics were observed. Implications for aetiology and clinical management are discussed.

Publication

Publisher: Springer Science and Business Media LLC

Date: 05-08-2007

DOI: 10.1007/S10989-007-9106-Z

Publication

Anticariogenic efficacy of a saliva biomimetic in head-and-neck cancer patients undergoing radiotherapy.

Publisher: Wiley

Date: 11-11-2019

DOI: 10.1111/ADJ.12658

Abstract: The aim of this study was to investigate the effect of treatment with the saliva biomimetic, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and SnF Twenty-four participants were randomized into two groups. Both groups used 0.4% SnF Both groups showed significantly reduced resting saliva flow rate (P < 0.001) postradiotherapy. There were no significant differences in flow rates and fluoride concentration between groups. The CPP-ACP group exhibited a significant (P < 0.05) 51% reduction in coronal surface caries progression compared with the placebo group. Resting salivary flow rate was significantly reduced in head-and-neck cancer patients following radiotherapy and use of CPP-ACP with SnF

Publication

Towards second-generation proteome analysis of murine enamel-forming cells

Publisher: Wiley

Date: 05-2006

DOI: 10.1111/J.1600-0722.2006.00310.X

Abstract: Proteome analysis of rat enamel-forming cells, initiated over a decade ago, has provided valuable insights to enamel biology. In preparation for a more comprehensive, second-generation proteomic exploration, we evaluated an updated micros le-profiling strategy that comprises sequential extraction of enamel epithelium, parallel one- and two-dimensional gel electrophoresis, and mass spectrometric sequence analysis. The results indicated that several hundred proteins, representing various cellular compartments (including membranes), are amenable to identification with a starting tissue volume of <10 microl. With its increased proteomic depth and breadth, this straightforward approach constitutes a major advance from the first-generation work (10-fold increased proteome coverage), although care was needed to ensure a comparably high stringency of protein identification. Expression proteomics has an exciting potential to elucidate the inner workings of murine enamel epithelial cells, leading to an improved understanding of enamel in health and disease.

Publication

Publisher: Wiley

Date: 12-2005

DOI: 10.1111/J.1834-7819.2005.TB00376.X

Publication

Date: 08-2005

DOI: 10.1111/J.1399-3011.2005.00273.X

Abstract: Bovine dentin phosphophoryn (BDP), a protein rich in aspartyl (Asp) and o-phosphoseryl [Ser(P)] residues, is synthesized by odontoblasts and believed to be involved in matrix-mediated biomineralization of dentin. The elucidation of the structure-function relationship of phosphophoryn has been a challenge because of its high-molecular weight, high negative charge, repetitive sequence, and lability. We have used the dynamic behavior of the (1)H NMR signal at 600 MHz to provide insight into the molecular dynamics of phosphophoryn. Our results indicate that phosphophoryn is a molecule of uniformly high mobility, thus belonging to a recently identified class of intrinsically disordered proteins that are characterized by sequences of low complexity and rich in polar and charged residues. The significance of our results is that phosphophoryn, because of its uniform nature has the potential to be replaced by biomimetic synthetic peptide analogs that together with amorphous calcium phosphate may lead to the development of novel, nontoxic, apatite-based dental restorative materials.

Publication

Polymerisation of a T cell epitope with an immunostimulatory C3d peptide sequence enhances antigen specific T cell responses

Publisher: Springer Science and Business Media LLC

Date: 03-2013

DOI: 10.1007/S10989-013-9343-2

Publication

Publisher: Microbiology Society

Date: 03-2010

DOI: 10.1099/MIC.0.033654-0

Abstract: Treponema denticola is an oral spirochaete that has been strongly associated with chronic periodontitis. The bacterium exists as part of a dense biofilm (subgingival dental plaque) accreted to the tooth. To determine T. denticola gene products important for persistence as a biofilm we developed a continuous-culture biofilm model and conducted a genome-wide transcriptomic analysis of biofilm and planktonic cells. A total of 126 genes were differentially expressed with a fold change of 1.5 or greater. This analysis identified the upregulation of putative prophage genes in the T. denticola 35405 genome. Intact bacteriophage particles were isolated from T. denticola and circular phage DNA was detected by PCR analysis. This represents the first, to our knowledge, functional bacteriophage isolated from T. denticola , which we have designated φtd1. In biofilm cells there was also an upregulation of genes encoding several virulence factors, toxin–antitoxin systems and a family of putative transposases. Together, these data indicate that there is a higher potential for genetic mobility in T. denticola when growing as a biofilm and that these systems are important for the biofilm persistence and therefore virulence of this bacterium.

Publication

Date: 06-03-2014

DOI: 10.1371/JOURNAL.PPAT.1003955

Publication

Porphyromonas gingivalis and the pathogenesis of Alzheimer’s disease

Publisher: Informa UK Limited

Date: 04-01-2023

DOI: 10.1080/1040841X.2022.2163613

Publication

The Porphyromonas gingivalis Ferric Uptake Regulator Orthologue Binds Hemin and Regulates Hemin-Responsive Biofilm Development

Publisher: Public Library of Science (PLoS)

Date: 06-11-2014

DOI: 10.1371/JOURNAL.PONE.0111168

Publication

Publisher: Wiley

Date: 12-10-2009

DOI: 10.1111/J.1399-302X.2009.00544.X

Publication

Vaccination with recombinant adhesins from the RgpA-Kgp proteinase-adhesin complex protects against Porphyromonas gingivalis infection.

Publisher: Elsevier BV

Date: 10-2006

DOI: 10.1016/J.VACCINE.2006.06.013

Abstract: Porphyromonas gingivalis is a pathogen associated with chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. A major virulence factor for P. gingivalis is the RgpA-Kgp proteinase-adhesin complex. We have prepared the following recombinant proteins corresponding to domains/regions of the RgpA-Kgp complex rRgpA(cat), rRgpA(A1), rRgpA(A2), rRgpA(A4), rRgpA(A1)(784-1009), rRgpA(A1)(911-1009), rKgp(A1) and rKgp(A1)(759-989). The ability of each recombinant protein to attenuate P. gingivalis infection, when used as a vaccine, in the murine lesion model was determined. All of the recombinant adhesin domains were found to significantly attenuate P. gingivalis infection with the most effective being rKgp(A1) and rKgp(A1)(759-989) followed by rRgpA(A1), rRgpA(A1)(784-1009) and rRgpA(A1)(911-1009). The predominant antibody isotype was IgG1. The A1 adhesins, which gave the best protection, contain specific motifs implicated in binding to host tissue. Immunisation with rRgpA(cat) had no effect on P. gingivalis infection. As well as detecting the Kgp(A1) adhesin in a Western blot, the rKgp(A1) and rKgp(A1)(759-989) antisera were also immunoreactive with the A1 and A3 adhesins of RgpA and HagA. Flow cytometric analysis indicated that rKgp(A1) and rRgpA(A1) antisera recognised native antigen on P. gingivalis whole cells. Furthermore, the rKgp(A1) and rKgp(A1)(759-989) antisera exhibited a similar immunoreactive profile with outer membrane preparations of all P. gingivalis serotypes and clinical isolates tested. The recombinant A1 adhesin therefore has potential in the development of a P. gingivalis vaccine.

Publication

Complementation in trans of Porphyromonas gingivalis Lipopolysaccharide Biosynthetic Mutants Demonstrates Lipopolysaccharide Exchange.

Publisher: American Society for Microbiology

Date: 21-04-2021

DOI: 10.1128/JB.00631-20

Abstract: Porphyromonas gingivalis is a keystone pathogen contributing to periodontitis in humans, leading to tooth loss. The oral microbiota is essential in this pathogenic process and changes from predominantly Gram-positive (health) to predominantly Gram-negative (disease) species. P. gingivalis uses its type IX secretion system (T9SS) to secrete and conjugate virulence proteins to anionic lipopolysaccharide (A-LPS).

Publication

Porphyromonas gingivalis Peptidylarginine Deiminase, a Key Contributor in the Pathogenesis of Experimental Periodontal Disease and Experimental Arthritis

Publisher: Public Library of Science (PLoS)

Date: 24-06-2014

DOI: 10.1371/JOURNAL.PONE.0100838

Publication

Publisher: Oxford University Press (OUP)

Date: 06-09-2013

DOI: 10.1111/1567-1364.12068

Publication

Publisher: Wiley

Date: 18-08-2008

DOI: 10.1111/J.1834-7819.2008.00061.X

Publication

Publisher: Public Library of Science (PLoS)

Date: 10-06-2013

DOI: 10.1371/JOURNAL.PONE.0065447

Publication

Publisher: Informa UK Limited

Date: 2013

DOI: 10.3402/JOM.V5I0.20320

Publication

Oxantel disrupts polymicrobial biofilm development of periodontal pathogens.

Publisher: American Society for Microbiology

Date: 2014

DOI: 10.1128/AAC.01375-13

Abstract: Bacterial pathogens commonly associated with chronic periodontitis are the spirochete Treponema denticola and the Gram-negative, proteolytic species Porphyromonas gingivalis and Tannerella forsythia . These species rely on complex anaerobic respiration of amino acids, and the anthelmintic drug oxantel has been shown to inhibit fumarate reductase (Frd) activity in some pathogenic bacteria and inhibit P. gingivalis homotypic biofilm formation. Here, we demonstrate that oxantel inhibited P. gingivalis Frd activity with a 50% inhibitory concentration (IC 50 ) of 2.2 μM and planktonic growth of T. forsythia with a MIC of 295 μM, but it had no effect on the growth of T. denticola . Oxantel treatment caused the downregulation of six P. gingivalis gene products and the upregulation of 22 gene products. All of these genes are part of a regulon controlled by heme availability. There was no large-scale change in the expression of genes encoding metabolic enzymes, indicating that P. gingivalis may be unable to overcome Frd inhibition. Oxantel disrupted the development of polymicrobial biofilms composed of P. gingivalis , T. forsythia , and T. denticola in a concentration-dependent manner. In these biofilms, all three species were inhibited to a similar degree, demonstrating the synergistic nature of biofilm formation by these species and the dependence of T. denticola on the other two species. In a murine alveolar bone loss model of periodontitis oxantel addition to the drinking water of P. gingivalis -infected mice reduced bone loss to the same level as the uninfected control.

Publication

Major proteins and antigens of Treponema denticola.

Publisher: Elsevier BV

Date: 10-2009

DOI: 10.1016/J.BBAPAP.2009.06.001

Abstract: Treponema denticola is a Gram-negative, motile, asaccharolytic, anaerobic spirochaete which along with Porphyromonas gingivalis and Tannerella forsythia has been shown to form a bacterial consortium called the Red Complex that is strongly associated with the clinical progression of chronic periodontitis. T. denticola was grown in continuous culture in a complex medium with a mean generation time of 15.75 h. S les from two different membrane-enriched preparations and a cytoplasm-enriched preparation were separated by two-dimensional gel electrophoresis and the proteins identified by MALDI-TOF/TOF mass spectrometry. In total, 219 non-redundant proteins were identified including numerous virulence factors, lipoproteins, ABC transporter proteins and enzymes involved in the metabolism of nine different amino acids of which glycine seems to be of particular importance. Novel findings include the identification of several abundant peptide uptake systems, and the identification of three flagellar filament outer layer proteins. Two-dimensional Western blot analysis using sera from mice immunized with formalin-killed T. denticola cells suggested that Msp, PrcA, OppA, OppA10, MglB, TmpC and several flagellar filament proteins are antigenic.

Publication

A Review of the Salivary Proteome and Peptidome and Saliva-derived Peptide Therapeutics

Publisher: Springer Science and Business Media LLC

Date: 06-09-2007

DOI: 10.1007/S10989-007-9109-9

Publication

Publisher: Informa UK Limited

Date: 2020

DOI: 10.1080/20002297.2020.1808750

Publication

Characterization of hemin-binding protein 35 (HBP35) in Porphyromonas gingivalis: Its cellular distribution, thioredoxin activity and role in heme utilization

Publisher: Springer Science and Business Media LLC

Date: 25-05-2010

DOI: 10.1186/1471-2180-10-152

Abstract: The periodontal pathogen Porphyromonas gingivalis is an obligate anaerobe that requires heme for growth. To understand its heme acquisition mechanism, we focused on a hemin-binding protein (HBP35 protein), possessing one thioredoxin-like motif and a conserved C-terminal domain, which are proposed to be involved in redox regulation and cell surface attachment, respectively. We observed that the hbp35 gene was transcribed as a 1.1-kb mRNA with subsequent translation resulting in three proteins with molecular masses of 40, 29 and 27 kDa in the cytoplasm, and one modified form of the 40-kDa protein on the cell surface. A recombinant 40-kDa HBP35 exhibited thioredoxin activity in vitro and mutation of the two putative active site cysteine residues abolished this activity. Both recombinant 40- and 27-kDa proteins had the ability to bind hemin, and growth of an hbp35 deletion mutant was substantially retarded under hemin-depleted conditions compared with growth of the wild type under the same conditions. P. gingivalis HBP35 exhibits thioredoxin and hemin-binding activities and is essential for growth in hemin-depleted conditions suggesting that the protein plays a significant role in hemin acquisition.

Publication

A therapeutic Porphyromonas gingivalis gingipain vaccine induces neutralising IgG1 antibodies that protect against experimental periodontitis

Publisher: Springer Science and Business Media LLC

Date: 12-2016

DOI: 10.1038/NPJVACCINES.2016.22

Abstract: Porphyromonas gingivalis infected mice with an established P. gingivalis -specific inflammatory immune response were protected from developing alveolar bone resorption by therapeutic vaccination with a chimera (KAS2-A1) immunogen targeting the major virulence factors of the bacterium, the gingipain proteinases. Protection was characterised by an antigen-specific IgG1 isotype antibody and Th2 cell response. Adoptive transfer of KAS2-A1-specific IgG1 or IgG2 expressing B cells confirmed that IgG1-mediated protection. Furthermore, parenteral or intraoral administration of KAS2-A1-specific polyclonal antibodies protected against the development of P. gingivalis -induced bone resorption. The KAS2-A1-specific antibodies neutralised the gingipains by inhibiting: proteolytic activity, binding to host cells roteins and co-aggregation with other periodontal bacteria. Combining key gingipain sequences into a chimera vaccine produced an effective therapeutic intervention that protected against P. gingivalis -induced periodontitis.

Publication

NMR studies of a novel calcium, phosphate and fluoride delivery vehicle-αS1-casein(59–79) by stabilized amorphous calcium fluoride phosphate nanocomplexes

Publisher: Elsevier BV

Date: 09-2004

DOI: 10.1016/J.BIOMATERIALS.2004.01.045

Publication

Kappacin, a novel antibacterial peptide from bovine milk.

Publisher: American Society for Microbiology

Date: 08-2019

DOI: 10.1128/AAC.45.8.2309-2315.2001

Abstract: Caseinomacropeptide (CMP) is a heterogeneous C-terminal fragment (residues 106 to 169) of bovine milk κ-casein composed of glycosylated and phosphorylated forms of different genetic variants. We have demonstrated that CMP has growth-inhibitory activity against the oral opportunistic pathogens Streptococcus mutans and Porphyromonas gingivalis and against Escherichia coli . CMP was fractionated using reversed-phase high-performance liquid chromatography (RP-HPLC), and each fraction was tested for activity against S. mutans in a 96-well-plate broth assay. Fractions were characterized by N-terminal sequence analysis and mass spectrometry. The active form of CMP was shown to be the nonglycosylated, phosphorylated κ-casein (residues 106 to 169) [κ-casein(106–169)], which we have designated kappacin. Endoproteinase Glu-C was used to hydrolyze CMP, and the generated peptides were separated using RP-HPLC and gel filtration-HPLC and then tested for activity against S. mutans . The peptide Ser( P ) 149 κ-casein-A(138–158) was the only peptide generated by endoproteinase Glu-C digestion that exhibited growth-inhibitory activity. Peptides corresponding to the sequences of the inhibitory peptide Ser( P ) 149 κ-casein-A(138–158) and its nonphosphorylated counterpart κ-casein-A(138–158) were chemically synthesized and tested for antibacterial activity. The synthetic Ser( P ) 149 κ-casein-A(138–158) displayed growth-inhibitory activity against S. mutans (MIC, 59 μg/ml [26 μM]). The nonphosphorylated peptide, however, did not inhibit growth at the concentrations tested, indicating that phosphorylation is essential for activity.

Eric Reynolds

Researcher

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Publications

Multifunctional Antimicrobial Polypeptide-Selenium Nanoparticles Combat Drug-Resistant Bacteria

Protease-activated receptor 2 has pivotal roles in cellular mechanisms involved in experimental periodontitis

Immediate Implant Placement Postextraction Without Flap Elevation

Synergistic virulence of Porphyromonas gingivalis and Treponema denticola in a murine periodontitis model.

Role of phosphorylated aminoacyl residues in generating atypical consensus sequences which are recognized by casein kinase-2 but not by casein kinase-1

Colonization of rat molar teeth by mutans streptococci with different salivary agglutination characteristics

Characterization and expression of a novel Porphyromonas gingivalis outer membrane protein, Omp28

Erosive potential of beverages sold in australian schools

Recharge and increase in hardness of GIC with CPP-ACP/F

Complete Nucleotide Sequence of a Gene prtR of Porphyromonas gingivalis W50 Encoding a 132-kDa Protein That Contains an Arginine-Specific Thiol Endopeptidase Domain and a Hemagglutinin Domain

An ELISA for a multiphosphorylated peptide, α(s1)-casein(59-79)

Effect of azithromycin on a red complex polymicrobial biofilm

C-Terminal Domain Residues Important for Secretion and Attachment of RgpB in Porphyromonas gingivalis

Effect of milk on caries incidence and bacterial composition of dental plaque in the rat

Peripheral T helper cell profiles during management of periodontitis

The synthesis of phosphopeptides via the Bpoc-based approach

Effect of ozone and Tooth Mousse TM on the efficacy of peroxide bleaching

Ion release from calcium and fluoride containing dental varnishes.

Characterization of the O-Glycoproteome of Tannerella forsythia

Host immune responses to Porphyromonas gingivalis antigens

Regulation of the 18-kDa heat shock protein in Mycobacterium ulcerans: An alpha-crystallin orthologue that promotes biofilm formation

An efficient method for enumerating oral spirochetes using flow cytometry.

Comparative study of the measurement of enamel demineralization and remineralization using transverse microradiography and electron probe microanalysis

Microbiome-targeted interventions for the control of oral–gut dysbiosis and chronic systemic inflammation

Interplay between Porphyromonas gingivalis and EGF signalling in the regulation of CXCL14.

Resin infiltration of developmentally hypomineralised enamel

Streptococcus mutans biofilm disruption by κ-casein glycopeptide.

Combined proteomic and transcriptomic interrogation of the venom gland of Conus geographus uncovers novel components and functional compartmentalization.

PG0026 Is the C-terminal Signal Peptidase of a Novel Secretion System of Porphyromonas gingivalis

A central role for the Hsp90·Cdc37 molecular chaperone module in interleukin-1 receptor-associated-kinase-dependent signaling by Toll-like receptors

Kgp and RgpB, but not RgpA, are important for Porphyromonas gingivalis virulence in the murine periodontitis model.

Combating multidrug-resistant Gram-negative bacteria with structurally nanoengineered antimicrobial peptide polymers

Cation-dependent structural features of β-casein-(1‒25)

Comparative transcriptomic analysis of Porphyromonas gingivalisbiofilm and planktonic cells

Oral health risk factors for bisphosphonate-associated jaw osteonecrosis.

Interferon Regulatory Factor 6 Differentially Regulates Toll-like Receptor 2-dependent Chemokine Gene Expression in Epithelial Cells

A 1H-NMR study of the casein phosphopeptide αs1-casein(59-79)

Complete Amino Acid Sequence and Comparative Molecular Modeling of HPR from Streptococcus mutans Ingbritt

Characterization of the O-Glycoproteome of Porphyromonas gingivalis

Divalent metal cations increase the activity of the antimicrobial peptide kappacin

FimR and FimS: Biofilm Formation and Gene Expression in Porphyromonas gingivalis

Extracellular proteomes of M-CSF (CSF-1) and GM-CSF-dependent macrophages

Porphyromonas gingivalis gingipains: The molecular teeth of a microbial vampire

Identification of a New Membrane-associated Protein That Influences Transport/Maturation of Gingipains and Adhesins of Porphyromonas gingivalis

Immunization with the RgpA-Kgp proteinase-adhesin complexes of Porphyromonas gingivalis protects against periodontal bone loss in the rat periodontitis model

Molecular modelling of the multiphosphorylated casein phosphopeptide alpha(S1)-casein(59-79) based on NMR constraints

Relationship between degree of casein hydrolysis and phosphopeptide release

A prospective clinical study of non-submerged immediate implants: Clinical outcomes and esthetic results

Effects of soy and bovine milk beverages on enamel mineral content in a randomized, double-blind in situ clinical study

Effect of CPP-ACP addition on physical properties of zinc oxide non-eugenol temporary cements

Porphyromonas gingivalis RgpA and Kgp proteinases and adhesins are C terminally processed by the carboxypeptidase CPG70

Effect of casein and whey-protein solutions on caries experience and feeding patterns of the rat

MEK‐ERK signaling diametrically controls the stimulation of IL‐23p19 and EBI3 expression in epithelial cells by IL‐36γ

Pancreatic beta cells are highly susceptible to oxidative and ER stresses during the development of diabetes.

IL-36γ regulates mediators of tissue homeostasis in epithelial cells

Antigens of bacteria associated with periodontitis.

Sodium ion-driven serine/threonine transport in Porphyromonas gingivalis.

Microbiome profiles of non-responding and responding paired periodontitis sites within the same participants following non-surgical treatment

Fluoride content of tank water in Australia

In-vitro study of the effect of casein phosphopeptide amorphous calcium fluoride phosphate on iatrogenic damage to enamel during orthodontic adhesive removal

Nitric Oxide on Extracorporeal Membrane Oxygenation in Neonates and Children (NECTAR Trial): Protocol for a Randomized Controlled Trial

Porphyromonas gingivalis RgpA-Kgp proteinase-adhesin complexes penetrate gingival tissue and induce proinflammatory cytokines or apoptosis in a concentration-dependent manner.

Metallocarboxypeptidase G (Porphyromonas gingivalis)

The Hydroxyapatite Binding Domains of Statherin: Influence of Size and Charge

Characterization of tryptic casein phosphopeptides prepared under industrially relevant conditions

Response of Porphyromonas gingivalis to Heme Limitation in Continuous Culture

Type IX secretion: the generation of bacterial cell surface coatings involved in virulence, gliding motility and the degradation of complex biopolymers

Antimicrobial peptides and their potential as oral therapeutic agents

Identification of a novel heterodimeric outer membrane protein of Porphyromonas gingivalis by two‐dimensional gel electrophoresis and peptide mass fingerprinting

Virulence factors of the oral spirochete Treponema denticola

Role of RgpA, RgpB, and Kgp proteinases in virulence of Porphyromonas gingivalis W50 in a murine lesion model.

Fluoride and casein phosphopeptide-amorphous calcium phosphate.

The A-chain of insulin is a hot-spot for CD4 ⁺ T cell epitopes in human type 1 diabetes