Characterization of the Binding of Urokinase-Type Plasminogen Activator to the Asialoglycoprotein Receptor

D. C. Rijken; M. E. van der Kaaden; E. Groeneveld; M. M. Barrett-Bergshoeff; Th. J. C. van Berkel; J. Kuiper

doi:10.1055/s-0037-1613158

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 88(01): 83-88
DOI: 10.1055/s-0037-1613158

Review Article

Schattauer GmbH

Characterization of the Binding of Urokinase-Type Plasminogen Activator to the Asialoglycoprotein Receptor

Authors

D. C. Rijken

¹Gaubius Laboratory, TNO Prevention and Health, Leiden, The Netherlands
M. E. van der Kaaden

²Division of Biopharmaceutics, Sylvius Laboratory, Leiden/Amsterdam Center for Drug Research University of Leiden, Leiden, The Netherlands
E. Groeneveld

¹Gaubius Laboratory, TNO Prevention and Health, Leiden, The Netherlands
M. M. Barrett-Bergshoeff

¹Gaubius Laboratory, TNO Prevention and Health, Leiden, The Netherlands
Th. J. C. van Berkel

²Division of Biopharmaceutics, Sylvius Laboratory, Leiden/Amsterdam Center for Drug Research University of Leiden, Leiden, The Netherlands
J. Kuiper

²Division of Biopharmaceutics, Sylvius Laboratory, Leiden/Amsterdam Center for Drug Research University of Leiden, Leiden, The Netherlands

Abstract

Summary

In order to study the role of the asialoglycoprotein receptor (ASGPr) in the rapid plasma clearance of urokinase-type plasminogen activator (u-PA), a microtiter plate binding assay was developed using ASGPr purified from rat liver extracts. Urinary two-chain u-PA bound to immobilized ASGPr in a saturable manner with an EC₅₀ of 0.2 µM. Binding was inhibited by rabbit antibodies against the ASGPr. In line with the known carbohydrate specificity of the ASGPr, GalNAc proved to be the most effective inhibitor from a series of monosaccharides, followed by Gal and Fuc, whereas GlcNAc was ineffective. The N-linked oligosaccharides of urinary u-PA do not terminate with the common Gal-GlcNAc element, but with a GalNAc-GlcNAc element which is partially sulfated. Sulfated forms of u-PA were separated from non-sulfated forms by using the lectin Wisteria floribunda agglutinin. Only the non-sulfated forms of u-PA (30% of the total) appeared to bind to the ASGPr. From different u-PA preparations used for thrombolytic therapy only urinary u-PA and u-PA produced by kidney cell cultures strongly bound to the ASGPr, whereas (recombinant) u-PA expressed in mouse myeloma cells, Chinese hamster ovary cells or E. coli scarcely bound to the receptor. It is concluded that u-PA bearing non-sulfated GalNAc-GlcNAc elements is specifically recognized by the ASGPr present on liver cells.

Keywords

Asialoglycoprotein receptor - plasma clearance - plasminogen activator - urokinase-type plasminogen activator - thrombolytic therapy

Full Text

References

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