Neutral proteinases from articular chondrocytes in culture. 2. Metal-dependent latent neutral proteoglycanase, and inhibitory activity

Sapolsky, Asher I.; Malemud, Charles J.; Norby, David P.; Moskowitz, Roland W.; Matsuta, Kunio; Howell, David S.

doi:10.1016/0005-2744(81)90257-6

Cited by 51 publications

(24 citation statements)

References 25 publications

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“…What is perhaps unexpected is the partially spontaneous activation of the enzyme. When metalloproteases have been detected in cultures of chondrocytcs, the activity has been noted to be mostly latent (32,331. Latent enzymes are frequently susceptible to activation by various serine proteases.…”

Section: Discussionmentioning

confidence: 99%

Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage

Martel‐Pelletier¹,

Pelletier²,

Cloutier³

et al. 1984

Arthritis & Rheumatism

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155

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Proteases have been postulated to account for the progressive disappearance of matrix proteoglycans in osteoarthritic (OA) cartilage. The digestion of endogenous proteoglycans by neutral proteases in human OA cartilage homogenates has been measured and compared with that of normal age-matched controls. Cartilage was obtained from 16 patients at the time of knee arthroplasty and from 7 accident victims. Tissue blocks were cut from the tibia1 plateau; part was used for histologic grading of the severity of OA and part was homogenized for the quantification of neutral metalloand serine protease activities, based on the release of digested products from endogenous proteoglycans. Total metalloprotease activity (latent plus active forms) was elevated 3-to 10-fold in all diseased cartilage. This elevation was already significant in mild disease, but was greatest in samples of moderate to severe disease. The active form of the enzyme was highest at the center of erosions and decreased in the margins of the plateau. The digestion of proteoglycans, as distinct from their mere release from the tissue, was demonstrated by chromatography on Sepharose-CL2B and by large pore electrophoresis. Serine protease activity on proteoglycans was much lower than that of metalloprotease. The mean activity was highest in mild disease and declined in the severe disease samples, but the difference between these 2 groups and the controls was not statistically significant. The results of this study are consistent with the hypothesis that the neutral metalloproteases of cartilage are involved in the degradation of proteoglycans in osteoarthritis.Although the etiologic mechanisms of osteoarthritis remain unknown, one fundamental finding is always the progressive degeneration and erosion of articular cartilage. In early stages of the disease there is a consistent loss of matrix proteoglycans, as evaluated by histochemical and biochemical means (1-5).Accompanying this loss is an increase in water content, as well as disturbances in the chemistry of the remaining proteoglycans (1,(6)(7)(8)(9)(10) In the present study we have measured the activity of neutral proteases in human OA cartilage by their action on endogenous proteoglycans using a micromethod modified from the procedure of Sapolsky et a1 (15). We havc demonstrated a significant elevation of neutral metalloprotease activity in OA specimens, particularly in the modcrate-severe stages of the disease. A second protease activity of the serine type appeared to be quantitatively less important. Serine activity tended to rise in OA, but the elevation was not statistically significant. MATERIALS AND METHODS Chemicals. The chemicals used were either AmericanChemical Society certified or the best grade commercially available. DNA calf thymus type I, chondroitin sulfate, and phenylmethylsulfonyl fluoride (PMS-F) were purchased from Sigma Chemical Co., St. Louis, MO; bovine trypsin inhibitor from Boehringer Mannheim, Indianapolis, IN; paminophenylmercuric acetate (APMA), cetylpyridinium chloride, and ...

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Section: Discussionmentioning

confidence: 99%

Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage

Martel‐Pelletier¹,

Pelletier²,

Cloutier³

et al. 1984

Arthritis & Rheumatism

Self Cite

155

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“…In the most recent reports from the Fell and Dingle groups (24,25), these investigators suggest that a factor from synovium induces chondrocytes to degrade their matrix, and thereby they support our proposition that degradation of the cartilage matrix may start within the cartilage. Indeed, most recently, we found that lapine articular chondrocytes in cell culture release metaldependent neutral proteoglycanase activity (7). Since the proteoglycanase occurs within the cartilage, it is logical to postulate that it may initiate and carry on the degradation of proteoglycan in osteoarthritis.…”

Section: Discussionmentioning

confidence: 99%

“…Sources include cultured synovial fibroblasts ( 2 ) , synovial fluid (3), skin fibroblasts (4), bone marrow macrophages ( 5 ) , bone explants (6), cultured chondrocytes from rabbit articular cartilage (7), and the involuting uterus (8).…”

Section: Further Characterization Of a Neutral Metalloprotease Isolatmentioning

confidence: 99%

“…Dingle (I 3) suggests that chondrocytes "under pathological stimulation may in fact be the principal agent of the degeneration of their surrounding matrix." Indeed, in a recent study (7), researchers demonstrate that cultured chondrocytes release into extracellular media active and latent neutral metalloproteases that are destructive to prOteOglyCanS.…”

Section: Further Characterization Of a Neutral Metalloprotease Isolatmentioning

confidence: 99%

“…Cultured chondrocytes, likewise, produced only small amounts of neutral metalloprotease, so that only partial purification has been possible (7). T h e present study concerns the further purification and additional characterization of the metal-dependent cartilage neutral proteoglycanase, which was obtained by processing large amounts of human postmortem tissue.…”

Section: Further Characterization Of a Neutral Metalloprotease Isolatmentioning

confidence: 99%

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Further characterization of a neutral metalloprotease isolated from human articular cartilage

Sapolsky

Howell

1982

Arthritis & Rheumatism

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The neutral metalloprotease extracted from 1,200 gm of human articular cartilage was purified 1,400‐ to 2,400‐fold by diethylaminoethyl‐ and carboxymethyl‐Sephadex chromatography. Disc electrophoresis and an isoelectric focusing method resolved the neutral enzyme activity into 4 bands. All bands had a similar amino acid analysis and a similar molecular weight by sodium dodecylsulfate electrophoresis and gel filtration: 24,000–27,000 daltons. The enzyme degraded proteoglycan subunit and proteoglycan aggregate to products with a sedimentation coefficient of 3S, but at low dilutions the enzyme produced 19.3S fragments. It is postulated that this protease may contribute to the development of osteoarthritis from within the cartilage matrix.

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Glycosaminoglycan Degradation

Kresse¹,

GlüSSL²

1987

Advances in Enzymology - And Related Areas of Molecular Biology

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Neutral proteinases from articular chondrocytes in culture. 2. Metal-dependent latent neutral proteoglycanase, and inhibitory activity

Cited by 51 publications

References 25 publications

Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage

Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage

Further characterization of a neutral metalloprotease isolated from human articular cartilage

Glycosaminoglycan Degradation

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