Evidence that the inhibition of cartilage proteoglycan breakdown by mannosamine is not mediated via inhibition of glycosylphosphatidylinositol anchor formation.
The effect of mannosamine, an inhibitor of glycosylphosphatidylinositol (GPI) anchor formation, on chondrocyte-mediated cartilage proteoglycan breakdown was investigated using cartilage explant cultures. Mannosamine inhibited interleukin 1alpha-, tumour necrosis factor alpha- and retinoic acid-stimulated proteoglycan release from bovine nasal and articular cartilage, and retinoic acid-stimulated proteoglycan release from human cartilage. Its effects on two GPI-anchored proteins [the urokinase receptor, which binds urokinase-type plasminogen activator (uPA) to cell surfaces, and alkaline phosphatase] were also studied using bovine chondrocytes. Enzyme histochemistry and zymography demonstrated cell-associated uPA-like serine proteinase activity and PA activity respectively which was not reduced by treatment of chondrocytes with mannosamine at concentrations effective at inhibiting cartilage proteoglycan breakdown. Similarly, the activity of cell-associated alkaline phosphatase was not reduced, except at mannosamine concentrations much higher than those used to inhibit proteoglycan breakdown. These results demonstrate that inhibition of proteoglycan breakdown by mannosamine is too potent to be explained by an effect on GPI-anchor formation.
Bryson
H
H
Division of Human Metabolism & Clinical Biochemistry, University of Sheffield Medical School, Beech Hill Road, Sheffield S10 2RX, U.K.
Buttle
D J
DJ
Kozaci
L D
LD
Johnatty
R N
RN
Bunning
R A
RA
D000368:Aged; D000369:Aged, 80 and over; D000469:Alkaline Phosphatase / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002356:Cartilage / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism; D046508:Culture Techniques; D005260:Female; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006595:Hexosamines / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D019344:Lactic Acid / Q000378:metabolism; D009300:Nasal Septum / Q000187:drug effects / Q000378:metabolism; D011509:Proteoglycans / Q000187:drug effects / Q000378:metabolism*; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
D017261:Glycosylphosphatidylinositols; D006595:Hexosamines; D007375:Interleukin-1; D011509:Proteoglycans; D014409:Tumor Necrosis Factor-alpha; C008322:mannosamine; D019344:Lactic Acid; D000469:Alkaline Phosphatase; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator