Thrombin reduces large heparan sulfate proteoglycan molecules in cultured vascular endothelial cell layers through inhibition of core protein synthesis.
We investigated the alteration of heparan sulfate proteoglycans induced by thrombin in cultured vascular endothelial cells. Heparan sulfate proteoglycans, which were metabolically labeled with [3H] glucosamine and [35S] sulfate, were isolated by DEAE-Sephacel ion-exchange chromatography and characterized by molecular sieve gel filtration. Core proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [35S] amino acids-labeled heparan sulfate proteoglycans after digestion with hepartinase. It was revealed that the high molecular weight subclass of heparan sulfate proteoglycans in the cell layer was markedly decreased by thrombin without changes of the hydrodynamic size of the molecules and the molecular weight of heparan sulfate chains. In addition, thrombin decreased the amount of large heparan sulfate proteoglycan core protein with a molecular weight of approximately 400 kDa, probably perlecan core, in the cell layer and the conditioned medium. The present data suggest that thrombin-induced decrease in the amount of heparan sulfate in vascular endothelial cell layer includes a reduction of the number of large heparan sulfate proteoglycan perlecan molecules through a suppression of the core protein synthesis.
Fujii
N
N
Department of Environmental Science, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Kaji
T
T
Akai
T
T
Koizumi
F
F
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002845:Chromatography; D004730:Endothelium, Vascular / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D011498:Protein Precursors / Q000096:biosynthesis; D013917:Thrombin / Q000494:pharmacology*
D019812:Heparan Sulfate Proteoglycans; D011498:Protein Precursors; D013917:Thrombin