Effect of tunicamycin on insulin binding and on proteoglycan synthesis and distribution in Swarm rat chondrosarcoma cell cultures.
Tunicamycin, an inhibitor of dolichol-diphospho-N-acetylglucosamine formation and hence an inhibitor of N-linked oligosaccharide biosynthesis, suppressed total proteoglycan synthesis by Swarm rat chondrosarcoma chondrocytes without affecting the size of the proteoglycan molecule, its secretion from the cell, or its ability to be retained in the extracellular matrix. In addition, tunicamycin did not substantially alter the ability of the chondrocytes to polymerize glycosaminoglycan onto an exogenous beta-D-xyloside acceptor. A secondary effect of tunicamycin suppression of proteoglycan synthesis was that a lesser amount of newly synthesized proteoglycan diffused from the extracellular matrix into the culture medium. The ability of exogenous hyaluronic acid and proteoglycan to increase the percentage of newly synthesized 35S-proteoglycan in the medium in tunicamycin-treated cultures indicates that matrix retention of 35S-proteoglycan is related to the total extracellular uronic acid content rather than to the presence or absence of mannose oligosaccharides bound to the proteoglycan molecule. These noncytotoxic concentrations of tunicamycin (33-333 ng/ml) decreased [3H]mannose incorporation to the same extent that they decreased total [35S]sulfate and [3H]serine incorporation, and caused the chondrocyte to synthesize and secrete a species of beta-hexosaminidase that was mannose-deficient as assessed by its failure to bind to concanavalin A. The additional finding of decreased insulin binding to tunicamycin-treated chondrosarcoma chondrocytes suggested that the inhibition of proteoglycan synthesis was due to diminution of receptors which respond to stimulatory hormones.
D000118:Acetylglucosaminidase / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D002813:Chondrosarcoma / Q000378:metabolism*; D002846:Chromatography, Affinity; D003208:Concanavalin A; D005944:Glucosamine / Q000031:analogs & derivatives*; D007328:Insulin / Q000378:metabolism*; D007700:Kinetics; D011509:Proteoglycans / Q000096:biosynthesis / Q000378:metabolism*; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism*; D012513:Sarcoma, Experimental / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology*
D007328:Insulin; D011509:Proteoglycans; D003208:Concanavalin A; D014415:Tunicamycin; D011972:Receptor, Insulin; D000118:Acetylglucosaminidase; D005944:Glucosamine