Ultrafiltration-based assay for heparanase activity

Analytical Biochemistry
Sachio TsuchidaMasaki Yanagishita

Abstract

Heparanase, a mammalian endoglycosidase that specifically cleaves heparan sulfate (HS), has been found in many tissues. Platelet, liver, and placenta have been abundant sources for the study of the enzyme. Notably, certain malignant cells also have been found to produce large amounts of the enzyme, the levels of which often correlate with their invasive and metastatic properties. To study roles of heparanase in various biological situations, a reliable method measuring the enzyme activity is indispensable. In the past, measurement of heparanase enzyme activity was done either by the detection of the degradation of fluorescent or radiolabeled HS chains by gel filtration procedures or by the use of radiolabeled substrate conjugated to solid matrices for the easy separation of degraded HS chains. A newly developed procedure, presented in this article, measures degradation of radiolabeled HS chains in the aqueous buffer by detecting their degradation products using an ultrafiltration device, the Centricon 30. This procedure has several advantages over previous assay procedures that involved tedious processing such as gel filtration chromatography of each sample or the preparation of substrate HS proteoglycans conjugated to a solid ...Continue Reading

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Jun 22, 2002·The Journal of Biological Chemistry·Katarzyna A Podyma-InoueMasaki Yanagishita
Jul 3, 2003·The Journal of Biological Chemistry·Feng GongJin-ping Li

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Citations

Feb 7, 2006·Biochemical and Biophysical Research Communications·Itay ShafatNeta Ilan
Sep 5, 2019·Bio-protocol·Jyothi C Sistla, Umesh R Desai
Sep 5, 2019··JC SistlaUmesh Desai

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