Collagenolytic activity in sonic extracts of Tannerella forsythia.
OBJECTIVE
The purpose of the present study was to characterize the collagenolytic activity in a sonicated extract of Tannerella forsythia and to investigate the activation of proMMMP-2 and -9 by the T. forsythia extract.


METHODS
The T. forsythia extract was incubated with type I collagen. The cleaved products were then analyzed by SDS-PAGE using the method of Laemmli. We studied the effects of cysteine, DTT, CaCl(2), and various proteinase inhibitors on collagenolytic activity. A HT1080 cell culture supernatant containing proMMP-2 and -9 was incubated with the T. forsythia extract and analyzed for the activation of proMMP-2 and -9 by gelatin zymography.


RESULTS
The T. forsythia extract degraded type I collagen. Cysteine increased the collagenolytic activity of the extract, and 5mM CaCl(2) was required for this activity. The collagenolytic activity of T. forsythia was inhibited by N-ethylmaleimide, iodoacetamide, iodoacetic acid, EDTA, and leupeptin, but not by PMSF, E-64, TLCK, or TPCK. When proMMP-2 and -9 were incubated with the T. forsythia extract, gelatinases with the relative molecular masses of MMP-2 and -9 were produced.


CONCLUSIONS
The present study suggests that T. forsythia extract is able to degrade type I collagen and activate proMMP-2 and -9.
Kawase
Norio
N
Department of Endodontics, Aichi-Gakuin University, Suemori-dori, Chikusa-ku, Nagoya, Japan.
Kishi
Jun-ichi
J
Nakamura
Hiroshi
H
Hayakawa
Taro
T
D001439:Bacteroides / Q000378:metabolism*; D002122:Calcium Chloride / Q000494:pharmacology; D002403:Cathepsins / Q000037:antagonists & inhibitors; D045744:Cell Line, Tumor; D002614:Chelating Agents / Q000494:pharmacology; D024042:Collagen Type I / Q000378:metabolism*; D003545:Cysteine / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D007460:Iodoacetamide / Q000494:pharmacology; D019807:Iodoacetic Acid / Q000494:pharmacology; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000187:drug effects; D020780:Matrix Metalloproteinase 9 / Q000187:drug effects / Q000378:metabolism; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
D002614:Chelating Agents; D024042:Collagen Type I; D015853:Cysteine Proteinase Inhibitors; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; D007976:Leupeptins; D011480:Protease Inhibitors; D015842:Serine Proteinase Inhibitors; D014107:Tosyllysine Chloromethyl Ketone; D014108:Tosylphenylalanyl Chloromethyl Ketone; D010664:Phenylmethylsulfonyl Fluoride; D004492:Edetic Acid; D002403:Cathepsins; D018093:Gelatinases; C120487:pro-matrix metalloproteinase 9; C069860:progelatinase; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D007930:Leucine; C032854:leupeptin; D003545:Cysteine; D002122:Calcium Chloride; D005033:Ethylmaleimide; C024974:E 64; D019807:Iodoacetic Acid; D007460:Iodoacetamide