{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T23:38:23Z","timestamp":1784763503766,"version":"3.55.0"},"reference-count":42,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2015,10,27]],"date-time":"2015-10-27T00:00:00Z","timestamp":1445904000000},"content-version":"vor","delay-in-days":26,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University of Tennessee College of Dentistry Alumni Endowment Fund"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clinical &amp; Exp Dental Res"],"published-print":{"date-parts":[[2015,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Locally applied therapeutic agents have become established in the treatment of periodontal disease. Inhibition of human metalloproteases by metal\u2010chelating antibiotics contributes to the utility of local therapy. Adding inhibitors of bacterial proteases might extend and improve local therapy. The periodontal pathogen <jats:italic>Porphyromonas gingivalis<\/jats:italic> (Pg) produces two extracellular cysteine proteases (gingipains Rgp and Kgp) that are virulence factors and contribute to destruction of oral tissues. Our aims were to compare efficacy of protease inhibitors against gingipains and evaluate bactericidal activity of the inhibitors. Protease activity was measured in fluorescent assays with specific Rgp and Kgp substrates. Bacterial viability was measured with <jats:italic>Bac<\/jats:italic>Light\u2122 (Invitrogen, Inc., Carlsbad, CA) reagents. Pairs of inhibitors of Rgp and Kgp, respectively, were leupeptin and cathepsin B inhibitor II, KYT\u20101 and KYT\u201036, and PPACK and Z\u2010FK\u2010ck. The cysteine\u2010protease inhibitor E64 was also tested. Rgp activity was higher than Kgp activity, and activity was higher in Pg 33277 and 49417 cell suspensions than in media. Concentrations required for 50% inhibition of Rgp in cell suspensions were 2\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, 2\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, 2\u2009\u00d7\u200910<jats:sup>\u22128<\/jats:sup>, and 5\u2009\u00d7\u200910<jats:sup>\u22125<\/jats:sup>\u2009M for KYT\u20101, PPACK, leupeptin, and E64, respectively. Concentrations required for 50% Kgp inhibition were 5\u2009\u00d7\u200910<jats:sup>\u221210<\/jats:sup>, 1\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, and 5\u2009\u00d7\u200910<jats:sup>\u22128<\/jats:sup>\u2009M for Z\u2010FK\u2010ck, KYT\u201036, and cathepsin B inhibitor II. E64 did not inhibit Kgp. Inhibition of Rgp could be accounted for by competition for binding between the arginine residue of the substrate and the guanidinobutane portion of E64. PPACK was the least selective, with a 10\u2010fold difference in concentrations that inhibited Rgp and Kgp. KYT\u20101 and Z\u2010FK\u2010ck inhibited both Rgp and Kgp, but inhibitory concentrations differed by 10,000\u2010fold. At up to 1\u2009\u00d7\u200910<jats:sup>\u22124<\/jats:sup>\u2009M, only Z\u2010FK\u2010ck was bactericidal. KYT\u20101 and KYT\u201036 were remarkably effective even when used in cell suspensions in which bacterial proteins could bind inhibitors or compete for binding to gingipains. These inhibitors might prove useful as an addition to locally applied therapeutic agents.<\/jats:p>","DOI":"10.1002\/cre2.4","type":"journal-article","created":{"date-parts":[[2015,10,27]],"date-time":"2015-10-27T05:22:38Z","timestamp":1445923358000},"page":"18-25","source":"Crossref","is-referenced-by-count":11,"title":["Inhibitors of bacterial protease enzymes for periodontal therapy"],"prefix":"10.1002","volume":"1","author":[{"given":"Kalid N.","family":"Hosn","sequence":"first","affiliation":[{"name":"Periodontology Department University of Tennessee Health Science Center  Memphis Tennessee USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary Margaret","family":"Jefferson","sequence":"additional","affiliation":[{"name":"Bioscience Research Department, College of Dentistry University of Tennessee Health Science Center  Memphis Tennessee USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlton","family":"Leding","sequence":"additional","affiliation":[{"name":"Bioscience Research Department, College of Dentistry University of Tennessee Health Science Center  Memphis Tennessee USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Solomon","family":"Shokouh\u2010Amiri","sequence":"additional","affiliation":[{"name":"Bioscience Research Department, College of Dentistry University of Tennessee Health Science Center  Memphis Tennessee USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edwin L.","family":"Thomas","sequence":"additional","affiliation":[{"name":"Bioscience Research Department, College of Dentistry University of Tennessee Health Science Center  Memphis Tennessee USA"},{"name":"Microbiology, Immunology &amp; Biochemistry Department University of Tennessee Health Science Center  Memphis Tennessee USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,10,27]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1177\/154405910608500502"},{"key":"e_1_2_7_3_1","first-page":"1223","article-title":"Roles for Arg\u2010 and Lys\u2010gingipains in the disruption of cytokine responses and loss of viability of human endothelial cells by Porphyromonas gingivalis infection","volume":"383","author":"Baba A.","year":"2002","journal-title":"J. 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