Increased degradation of carbohydrate deficient arterial basement membrane-like material from 2-deoxyglucose modified myomedial cell cultures.
The effect of 1.6 mM 2-deoxyglucose on the incorporation of [3H]leucine, [3H]glucosamine and [35S]sulphate into arterial basement membrane-like (BM) material was evaluated. BM-like material was isolated from the cell-matrix layer of cultured arterial smooth muscle cells by a sonication-differential centrifugation technique. 1.6 mM 2-deoxyglucose inhibited the 24 hr incorporation of [3H]glucosamine into BM-like material by 46% with a reduction in both [3H]glucosamine labelled glycopeptides and glycosaminoglycans. A marked decrease in [35S]sulphate incorporation (reduced by 80%) was demonstrated suggesting that 2-deoxyglucose may affect sulphatation of glycosaminoglycans. At 1.6 mM 2-deoxyglucose no effect on [3H]leucine incorporation was found. By gel filtration on Bio-Gel P6 a heterogeneous mixture of shortened glycopeptides was found after 2-deoxyglucose. The electrophoretic mobility of fibronectin and other glycoprotein components of BM-like material was increased. The stability of carbohydrate deficient BM-like material against removal/degradation was evaluated. A significantly increased removal of [3H]leucine from insufficiently glycosylated BM-like material was observed after a 24 hr chase period. The increased removal/degradation of BM-like material formed in the presence of 2-deoxyglucose was found to be a cellular dependent event.
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D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism; D001485:Basement Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D003837:Deoxy Sugars / Q000494:pharmacology*; D003847:Deoxyglucose / Q000494:pharmacology*; D006020:Glycopeptides / Q000378:metabolism; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000378:metabolism; D007700:Kinetics; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D011817:Rabbits
D003837:Deoxy Sugars; D006020:Glycopeptides; D006025:Glycosaminoglycans; D002738:Chloroquine; D003847:Deoxyglucose