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Papers In Press, published online ahead of print July 29, 2004
Department of Cancer and Vascular Biology, Technion-Israel, Haifa, Haifa 31096
Corresponding Author: vlodavsk{at}cc.huji.ac.il
Heparanase is a mammalian endoglycosidase that degrades heparan sulfate (HS) at specific intra-chain sites, activity that is strongly implicated in cell dissemination associated with metastasis and inflammation. In addition to its structural role in extracellular matrix (ECM) assembly and integrity, HS sequesters a multitude of polypeptides that reside in the ECM as a reservoir. A variety of growth factors, cytokines, chemokines and enzymes can be released by heparanase activity and profoundly affect cell and tissue function. Thus, heparanase bioavailability, accessibility and activity should be kept tightly regulated. We provide evidence that HS is not only a substrate for, but also a regulator of heparanase. Addition of heparin or xylosides to cell cultures resulted in a pronounced accumulation of heparanase in the culture medium, while sodium chlorate had no such effect. Moreover, cellular uptake of heparanase was markedly reduced in HS-deficient CHO 745 mutant cells, heparan sulfate proteoglycan-deficient colon cancer HT-29 cells, as well as in heparinase-treated cells. We have also studied heparanase biosynthetic route and found that the half life of the active enzyme, is approximately 30 h. This, and previous localization studies suggest that heparanase resides in the endosomal/lysosomal compartment for a relatively long period of time and is likely to play a role in the normal turnover of HS. Co-localization studies and cell fractionation following heparanase addition have identified syndecan family members as candidates molecules responsible for heparanase uptake, providing an efficient mechanism that limits extracellular accumulation and function of heparanase.
J. Biol. Chem, 10.1074/jbc.M402131200
Submitted on February 26, 2004
Revised on July 29, 2004
Accepted on July 29, 2004
Heparanase uptake is mediated by cell membrane heparan sulfate proteoglycans
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