Inhibition of Porphyromonas gingivalis proteinases (gingipains) by chlorhexidine: synergistic effect of Zn(II).
OBJECTIVE
Gingipains, proteolytic enzymes produced by the periodontal pathogen Porphyromonas gingivalis, are regarded as virulence factors in the pathogenesis of periodontitis. Inhibition of gingipain activity therefore may have therapeutic potential, and it has been suggested that chlorhexidine may inhibit the activities of these enzymes. The purposes of the present study were to examine systematically the inhibitory effects of chlorhexidine on three purified gingipains and to determine the effect of Zn(II) on chlorhexidine inhibition.


METHODS
The activities of lys-gingipain (Kgp) and two forms of arg-gingipain (RgpB and HRgpA) were measured in the presence of varying concentrations of chlorhexidine and with chlorhexidine supplemented with Zn(II). Inhibition constants (K(i)'s) were determined for chlorhexidine alone and in the presence of Zn(II). Fractional inhibitory constant indices were calculated to assess the synergy of the chlorhexidine-Zn(II) inhibition.


RESULTS
RgpB, HRgpA, and Kgp were all inhibited by chlorhexidine with K(i)'s in the micromolar range. For RgpB and HRgpA, the inhibitory effects of chlorhexidine were enhanced 3-30-fold by Zn(II). The chlorhexidine-Zn(II) interaction was synergistic for inhibition of HRgpA and RgpB. For Kgp, the effect of Zn(II) on chlorhexidine inhibition was antagonistic.


CONCLUSIONS
Chlorhexidine is an effective inhibitor of gingipains, and the inhibition of R-gingipains is enhanced by Zn(II). A mixture of chlorhexidine and Zn(II) may be useful as an adjunct in the treatment of periodontitis and in the post-treatment maintenance of periodontitis patients.
Cronan
C A
CA
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602-7229, USA.
Potempa
J
J
Travis
J
J
Mayo
J A
JA
D018829:Adhesins, Bacterial / Q000187:drug effects*; D000891:Anti-Infective Agents, Local / Q000008:administration & dosage / Q000494:pharmacology*; D002710:Chlorhexidine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000187:drug effects*; D015853:Cysteine Proteinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D004336:Drug Antagonism; D004357:Drug Synergism; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D015032:Zinc / Q000008:administration & dosage / Q000494:pharmacology*; D019345:Zinc Acetate / Q000008:administration & dosage / Q000494:pharmacology
D018829:Adhesins, Bacterial; D000891:Anti-Infective Agents, Local; D015853:Cysteine Proteinase Inhibitors; D000080867:Gingipain Cysteine Endopeptidases; D003546:Cysteine Endopeptidases; D019345:Zinc Acetate; D015032:Zinc; C010882:chlorhexidine gluconate; D002710:Chlorhexidine