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In this study, we compared the <jats:italic>ex vivo<\/jats:italic> PKP2 expression in periodontitis gingival biopsies to periodontitis\u2010free subjects and assessed the <jats:italic>in vitro<\/jats:italic> role of PKP2 in gingival epithelial barrier function and the mechanism by which <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic> modulates PKP2 expression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Material and methods<\/jats:title><jats:p>Using reverse transcription quantitative real\u2010time PCR (RT\u2010qPCR), we determined <jats:italic>PKP2<\/jats:italic>\u00a0mRNA expression levels in gingival biopsies collected from 11 periodontally healthy, 10 experimental gingivitis, and 10 chronic periodontitis subjects. PKP2 protein expression in gingival biopsies was detected by immunohistochemistry. We then challenged primary gingival epithelial cells with bacteria including <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic>, <jats:italic>Campylobacter rectus<\/jats:italic>, and various Toll\u2010like receptor agonists. Western blot and immunofluorescence staining were used to detect protein expression. Inhibitors blocking proteases pathways were tested for <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic>\u2010mediated PKP2 protein degradations. We also knocked down endogenous epithelial PKP2 using lentiviral short\u2010hairpin RNA (shRNA) and evaluated cell proliferation, spreading, and barrier function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Periodontitis gingival biopsies had approximately twofold less <jats:italic>PKP2<\/jats:italic> mRNA than did healthy controls (<jats:italic>p<\/jats:italic>\u00a0&lt;\u00a0.05). PKP2 protein was predominantly expressed in gingival epithelium. In primary gingival epithelial cells, <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic> challenge increased <jats:italic>PKP2<\/jats:italic> mRNA levels, while protein expression decreased, which suggests that <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic> has a protein degradation mechanism. Cysteine proteases inhibitors greatly attenuated <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic>\u2010mediated PKP2 protein degradation. Epithelial cells with deficient PKP2 exhibited inhibited cell proliferation and spreading and failed to form monolayers. Finally, <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic> impaired gingival epithelial barrier function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>PKP2 appears to be critical in maintaining gingival epithelial barrier function and is susceptible to degradation by cysteine proteases produced by <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic>. Our findings have identified a mechanism by which <jats:italic>P<\/jats:italic>.<jats:italic>gingivalis<\/jats:italic> impairs epithelial barrier function by promoting PKP2 degradation.<\/jats:p><\/jats:sec>","DOI":"10.1111\/jre.12918","type":"journal-article","created":{"date-parts":[[2021,8,9]],"date-time":"2021-08-09T07:17:03Z","timestamp":1628493423000},"page":"1046-1057","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Impaired function of epithelial plakophilin\u20102 is associated with periodontal disease"],"prefix":"10.1111","volume":"56","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3046-4338","authenticated-orcid":false,"given":"Ning","family":"Yu","sequence":"first","affiliation":[{"name":"The Forsyth Institute Cambridge Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinmei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Periodontics College of Dentistry University of Iowa Iowa City, Iowa USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sherill T.","family":"Phillips","sequence":"additional","affiliation":[{"name":"Center for Oral and Systemic Diseases School of Dentistry University of North Carolina at Chapel Hill Chapel Hill North Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steven","family":"Offenbacher","sequence":"additional","affiliation":[{"name":"Center for Oral and Systemic Diseases School of Dentistry University of North Carolina at Chapel Hill Chapel Hill North Carolina USA"},{"name":"Department of Periodontology School of Dentistry University of North Carolina at Chapel Hill Chapel Hill North Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3004-4023","authenticated-orcid":false,"given":"Shaoping","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Periodontics 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