title: The biologic action of single-chain choriogonadotropin is not dependent on the individual disulfide bonds of the beta subunit.
abstract: Disrupting disulfide loops in the human chorionic gonadotropin beta subunit (CGbeta) inhibits combination with the alpha subunit. Because the bioactivity requires a heterodimer, studies on the role of disulfide bonds on receptor binding/signal transduction have previously been precluded. To address this problem, we bypassed the assembly step and genetically fused CGbeta subunits bearing paired cysteine mutations to a wild-type alpha (WTalpha) subunit. The changes altered secretion of the single-chain mutants which parallel that seen for the CGbeta monomeric subunit. Despite conformational changes in CG disulfide bond mutants (assayed by gel electrophoresis and conformationally sensitive monoclonal antibodies), the variants bind to the lutropin/CG receptor and activated adenylate cyclase in vitro. The data show that the structural requirements for secretion and bioactivity are not the same. The results also suggest that the extensive native subunit interactions determined by the cystine bonds are not required for signal transduction. Moreover, these studies demonstrate that the single-chain model is an effective approach to structure-activity relationships of residues and structural domains associated with assembly of multisubunit ligands.
authors.affiliation: Department of Molecular Biology & Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: D
authors.initials: D
authors.lastname: Ben-Menahem
authors.forename: M
authors.initials: M
authors.lastname: Kudo
authors.forename: M R
authors.initials: MR
authors.lastname: Pixley
authors.forename: A
authors.initials: A
authors.lastname: Sato
authors.forename: N
authors.initials: N
authors.lastname: Suganuma
authors.forename: E
authors.initials: E
authors.lastname: Perlas
authors.forename: A J
authors.initials: AJ
authors.lastname: Hsueh
authors.forename: I
authors.initials: I
authors.lastname: Boime
mesh_terms: D006063:Chorionic Gonadotropin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D009154:Mutation; D011487:Protein Conformation
chemical_list: D006063:Chorionic Gonadotropin; D004220:Disulfides