title: The human follitropin alpha-subunit C terminus collaborates with a beta-subunit cystine noose and an alpha-subunit loop to assemble a receptor-binding domain competent for signal transduction.
abstract: FSH is a member of the pituitary/placental glycoprotein hormone family including luteinizing hormone, thyroid-stimulating hormone, and chorionic gonadotropin. These heterodimeric hormones share a common alpha-subunit and a highly homologous but distinct beta-subunit. The determinant loop of the FSH beta-subunit acts both as a specificity discriminator and as an essential receptor-binding site. The three-dimensional structure of hCG illustrates the proximity of the determinant loop to the carboxyl-terminal residues of the common alpha-subunit. Thus, site-directed mutagenesis was used to mak high-resolution substitutions at this carboxyl-terminal locus. The effects of those substitutions were studied. Twelve single mutations and one composite mutation were made of the region of hFSH alpha 74-92, each residue substituted by alanine. Side chain replacement in this region of FSH proved to be detrimental to binding. hFSH with mutations of either alpha S85A, alpha T86A, alpha K91A, or alpha S92A only retained 10% or less of the hFSH receptor-binding activity, while compared to these, mutants alpha H79A, alpha Y88A, and alpha Y89A retained slightly more binding activity. The single mutant alpha F74A and composite mutant alpha V76A/E77A binding activity was reduced to half of that of wild-type (WT) hFSH. In contrast, mutations of either alpha K75A, alpha T80A, alpha H83A, or alpha H90A did not adversely affect receptor binding, demonstrating the specificity of observed effects. The hFSH and mutant hormones were tested in an in vitro bioassay for stimulation of progesterone production. Those mutations that did not affect receptor binding (alpha K75A, alpha T80A, alpha H83A, and alpha H90A) did not affect signal transduction, measured by progesterone responses. After comparison of wild-type and mutant hFSH activities determined in radioreceptor assays (ID50) and in vitro bioassays (ED50), it became evident that signal transduction correlated with receptor binding.
authors.affiliation: Department of Biomedical Sciences, School of Public Health, State University of New York at Albany 12201, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Arnold
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Lindau-Shepard
authors.forename: M L
authors.initials: ML
authors.lastname: Losavio
authors.forename: M T
authors.initials: MT
authors.lastname: Patrascu
authors.forename: J A
authors.initials: JA
authors.lastname: Dias
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003553:Cystine / Q000378:metabolism* / Q000502:physiology; D019281:Dimerization; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011374:Progesterone / Q000096:biosynthesis; D017434:Protein Structure, Tertiary; D011962:Receptors, FSH / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D011962:Receptors, FSH; D003553:Cystine; D011374:Progesterone; D005640:Follicle Stimulating Hormone