Cellular source, activation and inhibition of dental plaque collagenase.
Dental plaque is the major aetiological factor in periodontal diseases and contains several proteolytic enzymes. The origin of these proteinases is, however, poorly studied. This study was undertaken to characterize collagenase present in dental plaque of adult periodontitis patients. Vertebrate-type rather than bacterial-derived collagenase activity was detected in extracts of both supra- and subgingival dental plaque extracts of adult periodontitis patients. Dental plaque collagenase was found to exist predominantly in autoactive form. Dental plaque collagenase from periodontally healthy individuals existed in latent from. Latent dental plaque collagenase from periodontitis lesions could be activated by a 95 kD chymotrypsin-like proteinase from Treponema denticola and human leukocyte cathepsin G but not by human plasmin. Incubation of purified latent leukocyte collagenase with whole cells of Fusobacterium nucleatum, Eubacterium saburreum, Prevotella buccae and Porphyromonas gingivalis, however, did not result to the activation of the enzyme. Doxycycline in vitro inhibited dental plaque collagenase with an IC50-value of 20 microM. Dental plaque collagenase degraded more efficiently type I and II collagens than type III collagen. Western-blot analysis with specific anti-human neutrophil collagenase-antibody revealed that both in supra- and subgingival dental plaque extracts dental plaque collagenase had undergone proteolytic conversion from an 80 kD proform to a 58 kD active form which is associated with catalytic autoactivity as measured by functional collagenase assay. This reflects proteolytic activation of leukocyte collagenase in dental plaque probably by other proteases derived from potent periodontopathogenic bacteria such as T. denticola or other PMN proteases such as cathepsin G.(ABSTRACT TRUNCATED AT 250 WORDS)
Sorsa
T
T
Department of Periodontology, University of Helsinki, Finland.
Ding
Y L
YL
Ingman
T
T
Salo
T
T
Westerlund
U
U
Haapasalo
M
M
Tschesche
H
H
Konttinen
Y T
YT
D000328:Adult; D015153:Blotting, Western; D002384:Catalysis; D056649:Cathepsin G; D002403:Cathepsins / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000502:physiology*; D003773:Dental Plaque / Q000201:enzymology* / Q000473:pathology*; D004318:Doxycycline / Q000494:pharmacology; D010450:Endopeptidases / Q000502:physiology; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism; D005051:Eubacterium / Q000201:enzymology; D005341:Fibrinolysin / Q000494:pharmacology; D016967:Fusobacterium nucleatum / Q000201:enzymology; D018093:Gelatinases / Q000502:physiology; D006801:Humans; D007962:Leukocytes / Q000201:enzymology; D061965:Matrix Metalloproteinase Inhibitors; D009504:Neutrophils / Q000201:enzymology; D010518:Periodontitis / Q000201:enzymology; D010519:Periodontium / Q000201:enzymology; D016966:Porphyromonas gingivalis / Q000201:enzymology; D018720:Prevotella / Q000201:enzymology; D012697:Serine Endopeptidases / Q000494:pharmacology; D014208:Treponema / Q000201:enzymology
D004792:Enzyme Precursors; D061965:Matrix Metalloproteinase Inhibitors; D003094:Collagen; D002403:Cathepsins; D010450:Endopeptidases; D012697:Serine Endopeptidases; C063172:clipsin; C534094:CTSG protein, human; D056649:Cathepsin G; D005341:Fibrinolysin; D017364:Collagenases; D018093:Gelatinases; D004318:Doxycycline