Pharmacological characterization of soluble human FSH receptor extracellular domain

Abstract

Follicle-stimulating hormone (FSH) is a member of the glycoprotein hormone family that regulates gametogenesis and steroidogenesis. Glycoprotein hormones signal through a unique class of G-protein-coupled receptors (GPCRs) that have a long extracellular domain (ECD), which is the primary site for hormone binding. The hFSHR-ECD was expressed in insect cells as a C-terminal, epitope-tagged protein resulting in production of soluble active receptor in the intracellular compartment and in the secreted culture medium. Coexpression of hFSHR-ECD with FSHβ or FSHα/β increased the secretion of the truncated receptor. Pharmacological studies to assess ligand-receptor interactions without the transmembrane domains showed higher affinity values (K _Ds) for ^[125l]hFSH using mammalian-expressed full-length receptor, secreted hFSHR-ECD, or secreted hFSHR-ECD coexpressed with FSHβ, whereas the K _D value for hFSHR-ECD coexpressed with FSHα/β subunits showed lower affinity. Competition of other glycoprotein hormones for secreted hFSHR-ECD coexpressed with FSHβ or mammalian full-length hFSHR resulted in similar binding profiles, indicating analogous pharmacology. Finally, we have demonstrated that a small molecule, suramin, which has been reported to interact with the mammalian full-length FSHR, competes for the binding of ^[1251]hFSH by interacting directly at the hFSHR-ECD.