The native 67-kilodalton minor fimbria of Porphyromonas gingivalis is a novel glycoprotein with DC-SIGN-targeting motifs

Amir E. Zeituni, William McCaig, Elizabeth Scisci, David G. Thanassi, Christopher W. Cutler

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

We recently reported that the oral mucosal pathogen Porphyromonas gingivalis, through its 67-kDa Mfa1 (minor) fimbria, targets the C-type lectin receptor DC-SIGN for invasion and persistence within human monocyte-derived dendritic cells (DCs). The DCs respond by inducing an immunosuppressive and Th2-biased CD4+ T-cell response. We have now purified the native minor fimbria by ion-exchange chromatography and sequenced the fimbria by tandem mass spectrometry (MS/MS), confirming its identity and revealing two putative N-glycosylation motifs as well as numerous putative O-glycosylation sites. We further show that the minor fimbria is glycosylated by ProQ staining and that glycosylation is partially removed by treatment with β(1-4)-galactosidase, but not by classic N- and O-linked deglycosidases. Further monosaccharide analysis by gas chromatography-mass spectrometry (GC-MS) confirmed that the minor fimbria contains the DC-SIGN-targeting carbohydrates fucose (1.35 nmol/mg), mannose (2.68 nmol/mg), N-acetylglucosamine (2.27 nmol/mg), and N-acetylgalactosamine (0.652 nmol/mg). Analysis by transmission electron microscopy revealed that the minor fimbria forms fibers approximately 200 nm in length that could be involved in targeting or cross-linking DC-SIGN. These findings shed further light on molecular mechanisms of invasion and immunosuppression by this unique mucosal pathogen.

Original languageEnglish (US)
Pages (from-to)4103-4110
Number of pages8
JournalJournal of Bacteriology
Volume192
Issue number16
DOIs
StatePublished - Aug 1 2010
Externally publishedYes

Fingerprint

Porphyromonas gingivalis
Dendritic Cells
Glycoproteins
Glycosylation
Galactosidases
C-Type Lectins
Acetylgalactosamine
Fucose
Acetylglucosamine
Monosaccharides
Ion Exchange Chromatography
Immunosuppressive Agents
Mannose
Tandem Mass Spectrometry
Transmission Electron Microscopy
Gas Chromatography-Mass Spectrometry
Immunosuppression
Monocytes
Carbohydrates
Staining and Labeling

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

The native 67-kilodalton minor fimbria of Porphyromonas gingivalis is a novel glycoprotein with DC-SIGN-targeting motifs. / Zeituni, Amir E.; McCaig, William; Scisci, Elizabeth; Thanassi, David G.; Cutler, Christopher W.

In: Journal of Bacteriology, Vol. 192, No. 16, 01.08.2010, p. 4103-4110.

Research output: Contribution to journalArticle

Zeituni, Amir E. ; McCaig, William ; Scisci, Elizabeth ; Thanassi, David G. ; Cutler, Christopher W. / The native 67-kilodalton minor fimbria of Porphyromonas gingivalis is a novel glycoprotein with DC-SIGN-targeting motifs. In: Journal of Bacteriology. 2010 ; Vol. 192, No. 16. pp. 4103-4110.
@article{c0fa9da2b28b4256a820bd88a47df07c,
title = "The native 67-kilodalton minor fimbria of Porphyromonas gingivalis is a novel glycoprotein with DC-SIGN-targeting motifs",
abstract = "We recently reported that the oral mucosal pathogen Porphyromonas gingivalis, through its 67-kDa Mfa1 (minor) fimbria, targets the C-type lectin receptor DC-SIGN for invasion and persistence within human monocyte-derived dendritic cells (DCs). The DCs respond by inducing an immunosuppressive and Th2-biased CD4+ T-cell response. We have now purified the native minor fimbria by ion-exchange chromatography and sequenced the fimbria by tandem mass spectrometry (MS/MS), confirming its identity and revealing two putative N-glycosylation motifs as well as numerous putative O-glycosylation sites. We further show that the minor fimbria is glycosylated by ProQ staining and that glycosylation is partially removed by treatment with β(1-4)-galactosidase, but not by classic N- and O-linked deglycosidases. Further monosaccharide analysis by gas chromatography-mass spectrometry (GC-MS) confirmed that the minor fimbria contains the DC-SIGN-targeting carbohydrates fucose (1.35 nmol/mg), mannose (2.68 nmol/mg), N-acetylglucosamine (2.27 nmol/mg), and N-acetylgalactosamine (0.652 nmol/mg). Analysis by transmission electron microscopy revealed that the minor fimbria forms fibers approximately 200 nm in length that could be involved in targeting or cross-linking DC-SIGN. These findings shed further light on molecular mechanisms of invasion and immunosuppression by this unique mucosal pathogen.",
author = "Zeituni, {Amir E.} and William McCaig and Elizabeth Scisci and Thanassi, {David G.} and Cutler, {Christopher W.}",
year = "2010",
month = "8",
day = "1",
doi = "10.1128/JB.00275-10",
language = "English (US)",
volume = "192",
pages = "4103--4110",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "16",

}

TY - JOUR

T1 - The native 67-kilodalton minor fimbria of Porphyromonas gingivalis is a novel glycoprotein with DC-SIGN-targeting motifs

AU - Zeituni, Amir E.

AU - McCaig, William

AU - Scisci, Elizabeth

AU - Thanassi, David G.

AU - Cutler, Christopher W.

PY - 2010/8/1

Y1 - 2010/8/1

N2 - We recently reported that the oral mucosal pathogen Porphyromonas gingivalis, through its 67-kDa Mfa1 (minor) fimbria, targets the C-type lectin receptor DC-SIGN for invasion and persistence within human monocyte-derived dendritic cells (DCs). The DCs respond by inducing an immunosuppressive and Th2-biased CD4+ T-cell response. We have now purified the native minor fimbria by ion-exchange chromatography and sequenced the fimbria by tandem mass spectrometry (MS/MS), confirming its identity and revealing two putative N-glycosylation motifs as well as numerous putative O-glycosylation sites. We further show that the minor fimbria is glycosylated by ProQ staining and that glycosylation is partially removed by treatment with β(1-4)-galactosidase, but not by classic N- and O-linked deglycosidases. Further monosaccharide analysis by gas chromatography-mass spectrometry (GC-MS) confirmed that the minor fimbria contains the DC-SIGN-targeting carbohydrates fucose (1.35 nmol/mg), mannose (2.68 nmol/mg), N-acetylglucosamine (2.27 nmol/mg), and N-acetylgalactosamine (0.652 nmol/mg). Analysis by transmission electron microscopy revealed that the minor fimbria forms fibers approximately 200 nm in length that could be involved in targeting or cross-linking DC-SIGN. These findings shed further light on molecular mechanisms of invasion and immunosuppression by this unique mucosal pathogen.

AB - We recently reported that the oral mucosal pathogen Porphyromonas gingivalis, through its 67-kDa Mfa1 (minor) fimbria, targets the C-type lectin receptor DC-SIGN for invasion and persistence within human monocyte-derived dendritic cells (DCs). The DCs respond by inducing an immunosuppressive and Th2-biased CD4+ T-cell response. We have now purified the native minor fimbria by ion-exchange chromatography and sequenced the fimbria by tandem mass spectrometry (MS/MS), confirming its identity and revealing two putative N-glycosylation motifs as well as numerous putative O-glycosylation sites. We further show that the minor fimbria is glycosylated by ProQ staining and that glycosylation is partially removed by treatment with β(1-4)-galactosidase, but not by classic N- and O-linked deglycosidases. Further monosaccharide analysis by gas chromatography-mass spectrometry (GC-MS) confirmed that the minor fimbria contains the DC-SIGN-targeting carbohydrates fucose (1.35 nmol/mg), mannose (2.68 nmol/mg), N-acetylglucosamine (2.27 nmol/mg), and N-acetylgalactosamine (0.652 nmol/mg). Analysis by transmission electron microscopy revealed that the minor fimbria forms fibers approximately 200 nm in length that could be involved in targeting or cross-linking DC-SIGN. These findings shed further light on molecular mechanisms of invasion and immunosuppression by this unique mucosal pathogen.

UR - http://www.scopus.com/inward/record.url?scp=77955939003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955939003&partnerID=8YFLogxK

U2 - 10.1128/JB.00275-10

DO - 10.1128/JB.00275-10

M3 - Article

C2 - 20562309

AN - SCOPUS:77955939003

VL - 192

SP - 4103

EP - 4110

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 16

ER -