title: Profile of glycosaminoglycan-degrading glycosidases and glycoside sulfatases secreted by human articular chondrocytes in homeostasis and inflammation.
abstract: OBJECTIVE
  To determine enzymatic activities of the 8 key glycosaminoglycan-degrading glycosidases and glycoside sulfatases in cultured human articular chondrocytes and in synovial fluid from patients with osteoarthritis.
  
  
  METHODS
  The following enzymes were analyzed: hexosaminidase and its isoenzyme A, N-acetyl-alpha-D-glucosaminidase, beta-galactosidase, beta-glucuronidase, alpha-L-iduronidase, aryl sulfatase, and galactose-6-sulfate sulfatase. Activity of the selected enzymes was analyzed by fluorometry with the aid of 4-methylumbelliferryl derivatives of the appropriate monosaccharides.
  
  
  RESULTS
  Hexosaminidase was found to be the dominant enzyme released by chondrocytes into the extracellular compartment. Stimulation of chondrocytes with interleukin-1beta resulted in a selective increase of the extracellular hexosaminidase activity and, to a lesser degree, of the extracellular beta-galactosidase activity, without significant changes in the activity of the other studied enzymes. Analysis of the pH dependency of the enzymatic activities revealed that even at neutral pH, hexosaminidase expressed a measurable activity, much higher than the activity of the other studied enzymes. Chondrocyte apoptosis did not result in increased extracellular glycosidase activities, including hexosaminidase activity. The spectrum of glycosidase and glycoside sulfatase activities in the synovial fluid from patients with osteoarthritis was similar to that in cultured human articular chondrocytes.
  
  
  CONCLUSIONS
  These data support the concept that lysosomal glycosidases, in particular hexosaminidase, represent a distinct subset of cartilage matrix-degrading enzymes that are activated by proinflammatory stimuli.
authors.affiliation: The Scripps Research Institute, and Scripps Clinic, La Jolla, California 92037, USA.
authors.forename: A R
authors.initials: AR
authors.lastname: Shikhman
authors.forename: D C
authors.initials: DC
authors.lastname: Brinson
authors.forename: M
authors.initials: M
authors.lastname: Lotz
mesh_terms: D017209:Apoptosis / Q000502:physiology; D002358:Cartilage, Articular / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000201:enzymology / Q000502:physiology; D005110:Extracellular Space / Q000201:enzymology; D006025:Glycosaminoglycans / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006027:Glycosides / Q000378:metabolism*; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018836:Inflammation Mediators / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D010003:Osteoarthritis / Q000201:enzymology / Q000473:pathology; D012016:Reference Values; D013429:Sulfatases / Q000378:metabolism*; D013582:Synovial Fluid / Q000201:enzymology; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D006025:Glycosaminoglycans; D006027:Glycosides; D018836:Inflammation Mediators; D007375:Interleukin-1; D013429:Sulfatases; D006026:Glycoside Hydrolases; D001619:beta-N-Acetylhexosaminidases