Structural Analysis of the Core Region of O-Lipopolysaccharide of
            <i>Porphyromonas gingivalis</i>
            from Mutants Defective in O-Antigen Ligase and O-Antigen Polymerase

Paramonov, Nikolay A.; Aduse‐Opoku, Joseph; Hashim, Ahmed; Rangarajan, Minnie; Curtis, Michael A.

doi:10.1128/jb.00019-09

Cited by 47 publications

(77 citation statements)

References 56 publications

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“…5C). The waaL transposon mutant lacks APS, which has been previously reported and serves as a control for the 1B5 antibody (20).…”

Section: Tn-seq Identification Of Pigment Genes In P Gingivalismentioning

confidence: 99%

“…Glycoprotein identification staining was performed using the Pierce glycoprotein staining kit (Invitrogen/Life Technologies) according to the manufacturer's instructions. For A-LPS Western blotting, nitrocellulose membranes were probed with monoclonal antibody (MAb) 1B5 directed against a glycan epitope on APS and arginine gingipain (20,38).…”

Section: Construction Of P Gingivalis Transposon Librariesmentioning

confidence: 99%

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Using Tn-seq To Identify Pigmentation-Related Genes of Porphyromonas gingivalis: Characterization of the Role of a Putative Glycosyltransferase

et al. 2017

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Cellular pigmentation is an important virulence factor of the oral pathogen Porphyromonas gingivalis. Pigmentation has been associated with many bacterial functions, including but not limited to colonization, maintaining a local anaerobic environment by binding oxygen molecules, and defense against reactive oxygen species (ROS) produced by immune cells. Pigmentation-associated loci identified to date have involved lipopolysaccharide, fimbriae, and heme acquisition and processing. We utilized a transposon mutant library of P. gingivalis strain ATCC 33277 and screened for pigmentation-defective colonies using massively parallel sequencing of the transposon junctions (Tn-seq) to identify genes involved in pigmentation. Transposon insertions at 235 separate sites, located in 67 genes and 15 intergenic regions, resulted in altered pigmentation: 7 of the genes had previously been shown to be involved in pigmentation, while 75 genes and intergenic regions had not. To further confirm identification, we generated a smaller transposon mutant library in P. gingivalis strain W83 and identified pigment mutations in several of the same loci as those identified in the screen in ATCC 33277 but also eight that were not identified in the ATCC 33277 screen. PGN_0361/ PG_0264, a putative glycosyltransferase gene located between two tRNA synthetase genes and adjacent to a miniature inverted-repeat transposable element, was identified in the Tn-seq screen and then verified through targeted deletion and complementation. Deletion mutations in PGN_0361/PG_0264 glycosyltransferase abolish pigmentation, modulate gingipain protease activity, and alter lipopolysaccharide. The mechanisms of involvement in pigmentation for other loci identified in this study remain to be determined, but our screen provides the most complete survey of genes involved in pigmentation to date. IMPORTANCE P. gingivalis has been implicated in the onset and progression of periodontal disease. One important virulence factor is the bacterium's ability to produce pigment. Using a transposon library, we were able to identify both known and novel genes involved in pigmentation of P. gingivalis. We identified a glycosyltransferase, previously not associated with pigmentation, that is required for pigmentation and determined its mechanism of involvement. A better understanding of the genes involved in pigmentation may lead to new insights into the complex mechanisms involved in this important virulence characteristic and could facilitate development of novel therapeutics.

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“…5C). The waaL transposon mutant lacks APS, which has been previously reported and serves as a control for the 1B5 antibody (20).…”

Section: Tn-seq Identification Of Pigment Genes In P Gingivalismentioning

confidence: 99%

Section: Construction Of P Gingivalis Transposon Librariesmentioning

confidence: 99%

Using Tn-seq To Identify Pigmentation-Related Genes of Porphyromonas gingivalis: Characterization of the Role of a Putative Glycosyltransferase

et al. 2017

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“…1B). Although the LPS structures of different strains of P. gingivalis may not be conserved (9,38,39,47), it is worth mentioning a study by Curtis et al in 1999 which characterized the LPS of P. gingivalis (9). This study determined that the LPS does not contain mannose or fucose, while our minor fimbria does.…”

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confidence: 99%

The Native 67-Kilodalton Minor Fimbria of Porphyromonas gingivalis Is a Novel Glycoprotein with DC-SIGN-Targeting Motifs

et al. 2010

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“…The inner core region lacks L-(D)-glycero-D(L)-manno-heptosyl residues and is linked to the outer core via 3-deoxy-D-mannooctulosonic acid (Kdo), which is attached to a glycerol (Gro) residue in the outer core via a monophosphodiester bridge (8). The outer region of the "uncapped core" is composed of a linear ␣-(1¡3)-linked D-Man OS with four or five members, one half of which are modified by phosphoethanolamine (PEA) at position O-6, which is attached to the glycerol at position O-2.…”

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confidence: 99%

Identification of the Linkage between A-Polysaccharide and the Core in the A-Lipopolysaccharide of Porphyromonas gingivalis W50

et al. 2015

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Porphyromonas gingivalis synthesizes two lipopolysaccharides (LPSs), O-LPS and A-LPS. The structure of the core oligosaccharide (OS) of O-LPS and the attachment site of the O-polysaccharide (O-PS) repeating unit [¡3)-␣-D-Galp-(1¡6)-␣-D-Glcp-(1¡4)-␣-L-Rhap-(1¡3)-␤-D-GalNAcp-(1¡ Porphyromonas gingivalis, a Gram-negative anaerobe, is frequently isolated from the subgingival plaque of periodontitis patients and is considered to be an important etiologic agent in periodontal disease. Among the major virulence factors of the organism are the cysteine proteases Arg gingipains (Rgps) and Lys gingipain (Kgp), specific for Arg-X and Lys-X peptide bonds, respectively, which are capable of degrading several host proteins (1), and lipopolysaccharide (LPS), which has the potential to alter the responses of the host innate defense system specifically through its lipid A component (2, 3).P. gingivalis W50 synthesizes two LPSs, namely O-LPS and A-LPS (4-6). Monoclonal antibody (MAb) 1B5, raised against one (RgpA cat ) of the isoforms of Arg gingipain, which are differentially glycosylated proteins, cross-reacts with A-LPS, suggesting that they share an epitope (7). Our group has identified the Man␣1-2Man␣1-phosphate fragment present in A-polysaccharide (A-PS) as part of the epitope recognized by MAb 1B5 (5). Thus, there appear to be common steps in the biosynthesis of A-LPS and the glycosylation of Arg gingipains in P. gingivalis.Analysis of an R-type LPS isolated from the P. gingivalis ⌬PG1051 mutant strain (waaL, encoding O-antigen ligase) enabled us to solve the structure of the core region of LPS (Fig. 1) (8).The inner core region lacks L-(D)-glycero-D(L)-manno-heptosyl residues and is linked to the outer core via 3-deoxy-D-mannooctulosonic acid (Kdo), which is attached to a glycerol (Gro) residue in the outer core via a monophosphodiester bridge (8). The outer region of the "uncapped core" is composed of a linear ␣-(1¡3)-linked D-Man OS with four or five members, one half of which are modified by phosphoethanolamine (PEA) at position O-6, which is attached to the glycerol at position O-2. In addition,

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Structural Analysis of the Core Region of O-Lipopolysaccharide of Porphyromonas gingivalis from Mutants Defective in O-Antigen Ligase and O-Antigen Polymerase

Cited by 47 publications

References 56 publications

Using Tn-seq To Identify Pigmentation-Related Genes of Porphyromonas gingivalis: Characterization of the Role of a Putative Glycosyltransferase

Using Tn-seq To Identify Pigmentation-Related Genes of Porphyromonas gingivalis: Characterization of the Role of a Putative Glycosyltransferase

The Native 67-Kilodalton Minor Fimbria of Porphyromonas gingivalis Is a Novel Glycoprotein with DC-SIGN-Targeting Motifs

Identification of the Linkage between A-Polysaccharide and the Core in the A-Lipopolysaccharide of Porphyromonas gingivalis W50

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