Molecular characteristics of extracellular matrix-associated glycosaminoglycans in endothelium

The extracellular matrix-associated sulfated glycosaminoglycans synthesized by bovine corneal endothelial cell clones consisted of 80% heparin sulfate and 20% chondroitin sulfate. The Mr distribution of haparan sulfate and chondroitin sulfte fractions were determined by high speed gel permeation chromatography. The average Mr of matrix-associated haparan sulfate of endothelial cell clones 1, 2,and3 were respectively 48000, 41 000, and 38 000. A size analysis of the degradation products after acidic nitrous acid deamination indicated that the degree of N-sulfation of heparin sulfate in clone 1 endothelial cells were relatively high. Heparan sulfate from all three clones was degraded by incubating purified haparan sulfate fractions with a metastatic B16 melanoma cell lysate containing heparanase (heparin sulfate specific endo-beta-glucuronidase). The average size of the heparin sulfate degradation products was Mr 10 000.