Activation of neutral metalloprotease in human osteoarthritic knee cartilage: evidence for degradation in the core protein of sulphated proteoglycan.
The neutral, metal dependent, proteoglycan degrading enzymes (NMPEs) in human osteoarthritic knee cartilage homogenates were activated by p-aminophenylmercuric acetate (APMA). The resultant effect on the structure of newly synthesised and already existing sulphated proteoglycan was measured. Newly synthesised and already existing proteoglycan aggregated to hyaluronic acid was reduced (p less than 0.01, p less than 0.05 respectively) when measured by chromatography on Sepharose CL-2B eluted with associative buffer. The APMA activated enzyme affected both the newly synthesised and already existing proteoglycan aggregate similarly (r = 0.79, p less than 0.001). Treatment of cartilage homogenates with APMA and 1,10-phenanthroline (10 mM) showed that the amount of aggregated proteoglycan was at the control level. The hydrodynamic size of the proteoglycan monomer (A1D1) was also reduced by treatment of cartilage homogenates with APMA. Reaggregation experiments with fraction A1D1 and exogenous hyaluronic acid and link protein showed a similar defect in forming proteoglycan aggregates. These data showed that activation of the NMPEs altered the structure of proteoglycan in two ways. The most consistent change was a reduction in the ability of proteoglycan to form aggregates with hyaluronic acid. This was likely to have occurred via a cleavage of the core protein in or around the hyaluronic acid binding globular domain. A second alteration probably includes a limited proteolytic cleavage in the remainder of the core protein.
Martel-Pelletier
J
J
Unité des Maladies Rhumatismales, Hôpital Notre-Dame, University of Montreal, Quebec, Canada.
Pelletier
J P
JP
Malemud
C J
CJ
D000368:Aged; D000369:Aged, 80 and over; D002358:Cartilage, Articular / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism; D007719:Knee Joint / Q000201:enzymology*; D046911:Macromolecular Substances; D008297:Male; D008666:Metalloendopeptidases / Q000378:metabolism*; D008875:Middle Aged; D009924:Organ Culture Techniques; D010003:Osteoarthritis / Q000201:enzymology*; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*
D046911:Macromolecular Substances; D011509:Proteoglycans; C010024:4-aminophenylmercuriacetate; D006820:Hyaluronic Acid; D008666:Metalloendopeptidases; D010662:Phenylmercuric Acetate