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Vol. 273, Issue 1, 544-551, January 2, 1998

The Inhibition of Fibroblast Growth Factor-2 Export by Cardenolides Implies a Novel Function for the Catalytic Subunit of Na+,K+-ATPase

Robert Z. Florkiewicz, Jerry Anchin, and Andrew Baird

From PRIZM Pharmaceuticals, San Diego, California 92121

Basic fibroblast growth factor (FGF-2) is one of a select group of proteins that can exit cells through an alternate, endoplasmic reticulum/Golgi apparatus independent exocytic pathway. This alternate pathway has been termed protein export. In an attempt to better understand this process, we have identified a family of related compounds, "cardenolides," that inhibit FGF-2 export. The cardenolides inhibit FGF-2 export in a time and concentration dependent fashion. Inhibition of FGF-2 export is specific in that the cardenolides have no effect on conventional protein secretion as measured by their inability to block release of the secreted protein human chorionic gonadotropin-alpha .

Because cardenolides are known to inhibit ion transport activity mediated by Na+,K+-ATPase, we investigated whether there are functional interactions between FGF-2 and their only known molecular target: the alpha -subunit of Na+,K+-ATPase. Export of FGF-2 from COS-1 cells is selectively inhibited when co-transfected with expression vectors encoding the alpha -subunit and FGF-2. Moreover, antibodies to the alpha -subunit specifically co-immunoprecipitate FGF-2 along with the alpha -subunit while conversely, antibodies to FGF-2 specifically co-immunoprecipitate the alpha -subunit along with FGF-2. Finally, the ion transporting activities of the Na+,K+-ATPase can be uncoupled from protein export. Varying the external concentration of K+ has little effect on export of FGF-2.

Taken together, these data: 1) identify a novel activity for cardenolides; 2) suggest a previously unknown role for the alpha -subunit of Na+, K+-ATPase in FGF-2 export; and 3) raise the possibility that the alpha -subunit itself may be an integral component of this alternate exocytic pathway mediating translocation of cytosolic FGF-2 to the cell surface.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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