Transglutaminase activity in aging articular chondrocytes and articular cartilage vesicles.
OBJECTIVE
Transglutaminases (TGases) (E.C. 2.3.2.13) catalyze a posttranslational modification of proteins and are associated with biomineralization in growth plate cartilage. Type II TGase participates in the activation of latent transforming growth factor beta (TGFbeta), a crucial factor for both normal cartilage mineralization and the pathologic mineralization that results in calcium pyrophosphate dihydrate (CPPD) crystal formation in aging articular cartilage. To explore a possible association between TGase levels and CPPD crystal formation in mature articular cartilage, TGase activity in articular chondrocytes from old and young pigs and in the articular cartilage vesicle (ACV) fraction of porcine articular cartilage was examined. In addition, the effects of TGase inhibitors on the production of inorganic pyrophosphate (PPi), a process necessary for CPPD crystallogenesis, were determined.


METHODS
TGase activity was measured with a radiometric assay in cultured articular chondrocytes from the knee joints of old (3-5 years old) and young (2-6 weeks old) pigs and in the ACVs. PPi levels were measured in chondrocyte-conditioned media in the presence of TGase inhibitors or control compounds.


RESULTS
Levels of TGase activity in the cytosolic fraction of old chondrocytes were 7-fold higher than those in identically cultured young chondrocytes. The mean +/- SD activity level in the membrane fraction of lysed chondrocytes was 6.0 +/- 0.6 units/mg protein in old articular chondrocytes and was undetectable in young chondrocytes. In ACVs, the mean +/- SD TGase activity level was 1.23 +/- 0.1 units/mg protein. Type II TGase protein was present in chondrocyte cytosol and in ACVs. TGase activity was increased by TGFbeta to 120% of control values (P < 0.01), and decreased by insulin-like growth factor 1 to 80% of control values (P < 0.01). TGase inhibitors blocked media accumulation of PPi, an essential precursor of CPPD crystal formation, and a sensitive marker of TGFbeta effect.


CONCLUSIONS
These data suggest a potential link between TGase activity and processes of pathologic biomineralization that result in CPPD crystal formation in aging articular cartilage.
Rosenthal
A K
AK
Medical College of Wisconsin, and the Zablocki VA Medical Center, Milwaukee 53295-1000, USA.
Derfus
B A
BA
Henry
L A
LA
D000375:Aging / Q000378:metabolism*; D000607:Aminoacetonitrile / Q000494:pharmacology; D000818:Animals; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002358:Cartilage, Articular / Q000166:cytology* / Q000201:enzymology*; D003538:Cystamine / Q000494:pharmacology; D011756:Diphosphates / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology; D006868:Hydrolysis / Q000187:drug effects; D008761:Methylhistamines / Q000494:pharmacology; D013552:Swine; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism*
D011756:Diphosphates; D004791:Enzyme Inhibitors; D006133:Growth Substances; D008761:Methylhistamines; D000607:Aminoacetonitrile; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine; D003538:Cystamine