PMID: 8590627Oct 1, 1995Paper

A competitive chromogenic assay to study the functional interaction of urokinase-type plasminogen activator with its receptor

Biological Chemistry Hoppe-Seyler
P RettenbergerM D Kramer

Abstract

Urokinase-type plasminogen activator (uPA) converts plasminogen to plasmin which degrades various extracellular matrix components. uPA is focused to the cell surface via binding to a specific receptor (uPAR, also termed CD87). uPAR-bound uPA mediates pericellular proteolysis in a variety of biological processes, e.g. cell migration, tissue remodeling and tumor invasion. We have developed a competitive microtiter plate-based chromogenic assay which allows the analysis of uPA/uPAR interaction. The plates are coated with recombinant uPAR expressed in Chinese hamster ovary (CHO) cells. Proteolytically active uPA (HMW-uPA) is added to the microtiter plate-attached uPAR. The amount of receptor-bound uPA is then determined indirectly via addition of plasminogen, which is activated to plasmin, followed by cleavage of a plasmin-specific chromogenic substrate. Substances interfering with binding of HMW-uPA to uPAR diminish the generation of plasmin, as indicated by a reduction of cleaved chromogenic substrate. This assay was used to analyze the inhibitory capacity of a variety of proteins and peptides, respectively, on the uPA/uPAR interaction: i) uPAR and uPAR-variants expressed in CHO cells, yeast or E. coli, ii) the aminoterminal frag...Continue Reading

References

Jul 1, 1992·Biological Chemistry Hoppe-Seyler·M SchmittH Graeff
Jul 1, 1995·Clinical & Experimental Metastasis·O WilhelmH Graeff
Feb 1, 1993·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·F Blasi

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Citations

Aug 28, 2012·Molecular Cancer·Bettina GrismayerViktor Magdolen
Jul 5, 2005·Expert Opinion on Investigational Drugs·U H Weidle, B König
Jul 29, 1997·International Journal of Cancer. Journal International Du Cancer·H de WitteK Danø

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