Collagenase, procollagenase and bone resorption effects of heparin, parathyroid hormone and calcitonin

Lenaers-Claeys, G; Vaes, Gilbert

doi:10.1016/0304-4165(79)90114-4

Cited by 50 publications

(17 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(ii) The rate of collagenase production from granuloma cells in primary culture was 1/100th of values reported for rheumatoid synovial cells in culture (8). (iii) Cell-mediated modulators produced %o to 5500th of enhanced enzyme production reported for bone explants (9) or rheumatoid synovial cells (8) in culture. (iv) The isoelectric point of granuloma collagenase (PI 6.2) is more acidic than values reported for rabbit tumor collagenase (PI 6.8 to7.0) (32).…”

Section: Discussionmentioning

confidence: 94%

Granuloma collagenase and EDTA-sensitive neutral protease production in hepatic murine schistosomiasis

Takahashi

Simpser

1981

Hepatology

View full text Add to dashboard Cite

Mice infected with Schistosoma mansoni represent a model for study of hepatic fibrosis in humans. Production of trypsin-activatable inactive collagenase and EDTA-sensitive neutral protease was measured in the culture medium in which granuloma explants or primary cultures were maintained. Collagenase production was maximal in granulomas obtained from liver of mice 8 weeks postinfection and was inhibited by Actinomycin D or cycloheximide, and enhanced by lymphocyte factor(s) or heparin. Isolated schistosome eggs did not release these enzymatic activities but did release EDTA-insensitive protease activity. Both enzymes were separated by ion-exchange chromatography and purified to homogeneity. Isolated collagenase had an isoelectric point of 6.2 and molecular weight of 60,000 and had the functional characteristics of a tissue collagenase. The specific activity of collagenase was 33 units per mg protein at an optimum pH 7.5 and lacked proteolytic activity against noncollagenous protein substrates. Isolated EDTA-sensitive neutral protease had specific caseinolytic activity of 150 units per mg protein and gelatinolytic activity of 300 units per mg protein at an optimum pH 7.5; the enzyme lacked activity against undenatured collagen. Isoelectric point was pH 6.0. Protease activity was inhibited by known inhibitors of collagenases. Production and activation of EDTA-sensitive neutral protease and collagenase accompany increased collagen synthesis and content in the liver of mice 8 weeks postinfection with S. mansoni cercariae. Continued accumulation of liver collagen under these conditions suggests an insufficiency in collagenase activity relative to the increase in collagen synthesis.

show abstract

Section: Discussionmentioning

confidence: 94%

Granuloma collagenase and EDTA-sensitive neutral protease production in hepatic murine schistosomiasis

Takahashi

Simpser

1981

Hepatology

View full text Add to dashboard Cite

show abstract

“…The bone explants released only insignificant amounts of either latent or active collagenase (Lenaers-Claeys & Vaes, 1976) or neutral proteinase (casein substrate) over 4 days' culture in a medium lacking heparin. The addition of increasing doses of heparin (12.5-600ig/ml) to the culture fluid had no effect on the accumulation of the active enzymes, but it caused a progressive accumulation of both procollagenase and latent neutral proteinase in the media (Fig.…”

Section: Resultsmentioning

confidence: 99%

The simultaneous release by bone explants in culture and the parallel activation of procollagenase and of a latent neutral proteinase that degrades cartilage proteoglycans and denatured collagen

Vaes¹,

Eeckhout²,

Lenaers-Claeys³

et al. 1978

Biochemical Journal

View full text Add to dashboard Cite

1. A latent neutral proteinase was found in culture media of mouse bone explants. Its accumulation during the cultures is closely parallel to that of procollagenase; both require the presence of heparin in the media. 2. Latent neutral proteinase was activated by several treatments of the media known to activate procollagenase, such as limited proteolysis by trypsin, chymotrypsin, plasmin or kallikrein, dialysis against 3 M-NaSCN at 4°C and prolonged preincubation at 25°C. Its activation often followed that of the procollagenase present in the same media. 3. Activation ofneutral proteinase (as does that ofprocollagenase) by trypsin or plasmin involved two successive steps: the activation of a latent endogenous activator present in the media followed by the activation of neutral proteinase itself by that activator. 4. The proteinase degrades cartilage proteoglycans, denatured collagen (Azocoll) and casein at neutral pH; it is inhibited by EDTA, cysteine or serum. Collagenase is not inhibited by casein or Azocoll and is less resistant to heat or to trypsin than is the proteinase. Partial separation of the two enzymes was achieved by gel filtration of the media but not by fractional (NH4)2SO4 precipitation, by ion exchange or by affinity chromatography on Sepharose-collagen. These fractionations did not activate latent enzymes. 5. Trypsin activation decreases the molecular weight of both latent enzymes

show abstract

“…A comparison of collagenolytic activity of these cathepsins has indicated that cathepsin L is particularly strong [4,13,14]. Vaes and his colleagues have shown that there is no correlation between the resorption of cultured mouse bone cells and their secretion of collagenase, while good correlations exist between bone resorption and the excretion of cathepsin B induced by parathyroid hormone (PTH) [6,15]. Furthermore, they also demonstrated that several inhibitors of cysteine proteinases, such as leupeptin, antipain, tosyl-lysyl chloromethane (Tos-Lys-CH,Cl), benzyl-oxycarbonylphenylalanyl-alanyl-diazomethane (Z-Phe-Ala-CHN,) and trans-epoxysuccinyl-L-leucylamido-(4-guanidino)-247 butane (E-64), markedly inhibited the resorption induced by PTH or heparin in cultured mouse bone cells [4-61.…”

Section: Introductionmentioning

confidence: 99%

Participation of cathepsin L on bone resorption

et al. 1993

View full text Add to dashboard Cite

The proteinase responsible for bone collagen degradation in osteo-resorption was examined. The bone pit formation induced by parathyroid hormone (PTH) was markedly suppressed by leupeptin, E-64 and cystatin A, while no inhibition was observed by CA-074, a specific inhibitor of cathepsin B. Pig leucocyte cysteine proteinase inhibitor (PLCPI), a specific inhibitor of cathepsin L, and chymostatin, a selective inhibitor of cathepsin L, completely inhibited the pit formation. Cathepsin L activity in osteoclasts was much higher than the other cathepsin activities. Serum calcium in rats placed on a low calcium diet was decreased by treatment of E-64 or cystatin A, but not by CA-074. These findings suggest that cathepsin L is the main proteinase responsible for bone collagen degradation.

show abstract

Collagenase, procollagenase and bone resorption effects of heparin, parathyroid hormone and calcitonin

Cited by 50 publications

References 22 publications

Granuloma collagenase and EDTA-sensitive neutral protease production in hepatic murine schistosomiasis

Granuloma collagenase and EDTA-sensitive neutral protease production in hepatic murine schistosomiasis

The simultaneous release by bone explants in culture and the parallel activation of procollagenase and of a latent neutral proteinase that degrades cartilage proteoglycans and denatured collagen

Participation of cathepsin L on bone resorption

Contact Info

Product

Resources

About