By using the yeast two-hybrid system, muskelin was found to bind with the carboxy-terminal tail of the prostaglandin EP3 receptor alpha isoform but not with either the beta or gamma isoform. A direct interaction between the carboxy-terminal tail of the alpha isoform and muskelin was confirmed in vitro using recombinant fusion proteins. Analysis by confocal microscopy indicated that the isoform and muskelin were distributed at the plasma membrane in transfected cells. When the isoform was stimulated by agonist, the receptor was internalized in the cells expressing the receptor alone, but this internalization was partially inhibited by the cotransfection with muskelin. Furthermore, muskelin enhanced the Gi activity of the isoform. Thus, muskelin appears to be an isoform-specific anchoring protein for the EP3 receptor.
Copyright 2000 Academic Press.