A synthetic peptide corresponding to glycoprotein hormone alpha subunit residues 32-46 inhibits gonadotropin binding to receptor.

Abstract

A synthetic peptide strategy was used to study structure-function relationships between residues 32 to 46 of the glycoprotein hormone alpha subunit (GPH alpha) and the testicular follicle-stimulating hormone (FSH) and luteinizing hormone (LH/hCG) receptors. A peptide amide corresponding to this region [GPH-alpha-(32-46)] inhibited both 125I-hFSH and 125I-hCG binding to their respective calf testis membrane receptors. The concentration at which GPH-alpha-(32-46) peptide amide inhibited FSH binding by 50% (IC50) was 36 microM, and for hCG it was 54 microM. GPH-alpha-(32-46) peptide amide also inhibited FSH-stimulated estradiol biosynthesis in cultured rat Sertoli cells. In order to determine the involvement of individual residues within this region of the glycoprotein hormone alpha subunit in receptor binding inhibitory activity, truncated and alanine-substituted peptide analogs were synthesized and tested in both FSH and hCG radioligand receptor competition assays. Based on the relative potency of each peptide, we conclude that Phe-33, Arg-35, Arg-42, Ser-43 and Lys-44 may be important, and Cys-32 is required, for inhibition of FSH and hCG binding to their respective receptor. Our results demonstrate involvement of the glycoprotein hormone alpha-subunit in receptor binding, identify residues 32 to 46 as a receptor binding domain, and define the relative importance of specific residues within this region of the alpha subunit for hormone-receptor interaction.