Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes: A strategy for cell adhesion and migration disabling

Edith Hintermann, Susan Kinder Haake, Urs Christen, Andrew Sharabi, Vito Quaranta

Research output: Contribution to journalArticle

Abstract

Adhesive interactions of cells are critical to tissue integrity. We show that infection with Porphyromonas gingivalis, a major pathogen in the periodontal disease periodontitis, interferes with both cell-matrix and cell-cell adhesion in the oral keratinocyte cell line HOK-16. Thus, infected cells showed reduced adhesion to extracellular matrix, changes in morphology from spread to rounded, and impaired motility on purified matrices in Transwell migration assays and scratch assays. Western blot analysis of P. gingivalis-challenged HOK-16 cells revealed proteolysis of focal contact components (e.g., focal adhesion kinase), adherens junction proteins (e.g., catenins), and adhesion signaling molecules (e.g., the tyrosine kinase SRC). Proteolysis was selective, since important components of adherens junctions (E-cadherin) or signaling molecules (extracellular signal-regulated kinases ERK1/2) were not degraded. The virulence factors gingipains, cysteine proteinases expressed by P. gingivalis, are likely responsible for this proteolytic attack, since they directly digested specific proteins in pull-down experiments, and their proteolytic activity was blocked by the cysteine proteinase inhibitor N-a-p-tosyl-L-lysine chloromethyl ketone and also by a caspase inhibitor. Proteolysis was strain dependent, such that ATCC 33277 and 381 had high proteolytic potential, whereas W50 showed almost no proteolytic activity. These findings may help explain the formation of gingival pockets between cementum and periodontal epithelium, a hallmark of periodontitis. Furthermore, they illustrate a new pathogenetic paradigm of infection whereby bacteria may disrupt the integrity of epithelia.

Original languageEnglish (US)
Pages (from-to)5846-5856
Number of pages11
JournalInfection and Immunity
Volume70
Issue number10
DOIs
StatePublished - Oct 2002
Externally publishedYes

Fingerprint

Adherens Junctions
Porphyromonas gingivalis
Focal Adhesions
Keratinocytes
Cell Adhesion
Proteolysis
Cell Movement
Periodontitis
Gingival Pocket
Epithelium
Cysteine Proteinase Inhibitors
Dental Cementum
Catenins
Focal Adhesion Protein-Tyrosine Kinases
Caspase Inhibitors
Cysteine Proteases
Mitogen-Activated Protein Kinase 1
Periodontal Diseases
Virulence Factors
Cadherins

ASJC Scopus subject areas

  • Immunology

Cite this

Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes : A strategy for cell adhesion and migration disabling. / Hintermann, Edith; Haake, Susan Kinder; Christen, Urs; Sharabi, Andrew; Quaranta, Vito.

In: Infection and Immunity, Vol. 70, No. 10, 10.2002, p. 5846-5856.

Research output: Contribution to journalArticle

Hintermann, Edith ; Haake, Susan Kinder ; Christen, Urs ; Sharabi, Andrew ; Quaranta, Vito. / Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes : A strategy for cell adhesion and migration disabling. In: Infection and Immunity. 2002 ; Vol. 70, No. 10. pp. 5846-5856.
@article{1e32960e66114106ba9faf1aeb97589d,
title = "Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes: A strategy for cell adhesion and migration disabling",
abstract = "Adhesive interactions of cells are critical to tissue integrity. We show that infection with Porphyromonas gingivalis, a major pathogen in the periodontal disease periodontitis, interferes with both cell-matrix and cell-cell adhesion in the oral keratinocyte cell line HOK-16. Thus, infected cells showed reduced adhesion to extracellular matrix, changes in morphology from spread to rounded, and impaired motility on purified matrices in Transwell migration assays and scratch assays. Western blot analysis of P. gingivalis-challenged HOK-16 cells revealed proteolysis of focal contact components (e.g., focal adhesion kinase), adherens junction proteins (e.g., catenins), and adhesion signaling molecules (e.g., the tyrosine kinase SRC). Proteolysis was selective, since important components of adherens junctions (E-cadherin) or signaling molecules (extracellular signal-regulated kinases ERK1/2) were not degraded. The virulence factors gingipains, cysteine proteinases expressed by P. gingivalis, are likely responsible for this proteolytic attack, since they directly digested specific proteins in pull-down experiments, and their proteolytic activity was blocked by the cysteine proteinase inhibitor N-a-p-tosyl-L-lysine chloromethyl ketone and also by a caspase inhibitor. Proteolysis was strain dependent, such that ATCC 33277 and 381 had high proteolytic potential, whereas W50 showed almost no proteolytic activity. These findings may help explain the formation of gingival pockets between cementum and periodontal epithelium, a hallmark of periodontitis. Furthermore, they illustrate a new pathogenetic paradigm of infection whereby bacteria may disrupt the integrity of epithelia.",
author = "Edith Hintermann and Haake, {Susan Kinder} and Urs Christen and Andrew Sharabi and Vito Quaranta",
year = "2002",
month = "10",
doi = "10.1128/IAI.70.10.5846-5856.2002",
language = "English (US)",
volume = "70",
pages = "5846--5856",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "10",

}

TY - JOUR

T1 - Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes

T2 - A strategy for cell adhesion and migration disabling

AU - Hintermann, Edith

AU - Haake, Susan Kinder

AU - Christen, Urs

AU - Sharabi, Andrew

AU - Quaranta, Vito

PY - 2002/10

Y1 - 2002/10

N2 - Adhesive interactions of cells are critical to tissue integrity. We show that infection with Porphyromonas gingivalis, a major pathogen in the periodontal disease periodontitis, interferes with both cell-matrix and cell-cell adhesion in the oral keratinocyte cell line HOK-16. Thus, infected cells showed reduced adhesion to extracellular matrix, changes in morphology from spread to rounded, and impaired motility on purified matrices in Transwell migration assays and scratch assays. Western blot analysis of P. gingivalis-challenged HOK-16 cells revealed proteolysis of focal contact components (e.g., focal adhesion kinase), adherens junction proteins (e.g., catenins), and adhesion signaling molecules (e.g., the tyrosine kinase SRC). Proteolysis was selective, since important components of adherens junctions (E-cadherin) or signaling molecules (extracellular signal-regulated kinases ERK1/2) were not degraded. The virulence factors gingipains, cysteine proteinases expressed by P. gingivalis, are likely responsible for this proteolytic attack, since they directly digested specific proteins in pull-down experiments, and their proteolytic activity was blocked by the cysteine proteinase inhibitor N-a-p-tosyl-L-lysine chloromethyl ketone and also by a caspase inhibitor. Proteolysis was strain dependent, such that ATCC 33277 and 381 had high proteolytic potential, whereas W50 showed almost no proteolytic activity. These findings may help explain the formation of gingival pockets between cementum and periodontal epithelium, a hallmark of periodontitis. Furthermore, they illustrate a new pathogenetic paradigm of infection whereby bacteria may disrupt the integrity of epithelia.

AB - Adhesive interactions of cells are critical to tissue integrity. We show that infection with Porphyromonas gingivalis, a major pathogen in the periodontal disease periodontitis, interferes with both cell-matrix and cell-cell adhesion in the oral keratinocyte cell line HOK-16. Thus, infected cells showed reduced adhesion to extracellular matrix, changes in morphology from spread to rounded, and impaired motility on purified matrices in Transwell migration assays and scratch assays. Western blot analysis of P. gingivalis-challenged HOK-16 cells revealed proteolysis of focal contact components (e.g., focal adhesion kinase), adherens junction proteins (e.g., catenins), and adhesion signaling molecules (e.g., the tyrosine kinase SRC). Proteolysis was selective, since important components of adherens junctions (E-cadherin) or signaling molecules (extracellular signal-regulated kinases ERK1/2) were not degraded. The virulence factors gingipains, cysteine proteinases expressed by P. gingivalis, are likely responsible for this proteolytic attack, since they directly digested specific proteins in pull-down experiments, and their proteolytic activity was blocked by the cysteine proteinase inhibitor N-a-p-tosyl-L-lysine chloromethyl ketone and also by a caspase inhibitor. Proteolysis was strain dependent, such that ATCC 33277 and 381 had high proteolytic potential, whereas W50 showed almost no proteolytic activity. These findings may help explain the formation of gingival pockets between cementum and periodontal epithelium, a hallmark of periodontitis. Furthermore, they illustrate a new pathogenetic paradigm of infection whereby bacteria may disrupt the integrity of epithelia.

UR - http://www.scopus.com/inward/record.url?scp=0036784687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036784687&partnerID=8YFLogxK

U2 - 10.1128/IAI.70.10.5846-5856.2002

DO - 10.1128/IAI.70.10.5846-5856.2002

M3 - Article

C2 - 12228316

AN - SCOPUS:0036784687

VL - 70

SP - 5846

EP - 5856

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 10

ER -