Functionally distinct agonist and receptor-binding regions in human chorionic gonadotropin. Development of a tertiary structure model.
The histidine residues in human chorionic gonadotropin (hCG) were chemically modified using diethyl pyrocarbonate. Derivatives of hCG with an average of 0.5-3.5 histidines modified (maximum of 4 per hCG) had reduced receptor-binding and cell-stimulating activities. Acylation of hCG at progressively lower pH values (conditions in which 1 of the 2 absolutely conserved histidines alpha His-83 is not titratable, whereas alpha His-94 becomes increasingly protonated and resistant to modification) produced hCG derivatives with a greater retention of receptor-binding activity than cell-stimulating activity. The involvement of alpha His-94 as part of the receptor-binding region of the hormone and of alpha His-83 as a putative active site residue was inferred. Proteinaceous protease inhibitors were shown to neutralize the agonist activity of hCG and to reduce the binding of hCG to its receptor and also to specific antisera. It was presumed that an inhibitor-hormone complex was formed which was analogous to the complexing of inhibitor with the "substrate pocket" of a serine protease. The discovery of primary sequence analogies between hCG and the serine protease chymotrypsin enabled the prediction of hCG structure using the enzyme as a folding template. Solvent-exposed and buried core regions of the peptide chain were delineated using smoothed hydrophobicity profiles in combination with Chou-Fasman secondary structure predictions. Hypervariable hydrophobicity indices between residues 38 and 80 of the human beta subunits reflected different folding arrangements which presumably conferred the individual receptor specificities. When mapped to the putative structure these receptor-determinant loops were adjacent to an area of the alpha subunit analogous to the substrate pocket of serine proteases. Disulfide bond assignments and intersubunit contact regions were identifiable. The proposed tertiary structure for hCG manifests the topographical epitopes defined using monoclonal antibodies and satisfies the currently available data on specific modification and its effects upon hormonal structure and function. This paper is considered to be the first report of a differential effect upon the agonist and receptor binding abilities of a glycoprotein hormone after modification of the proteinaceous, as opposed to the glycosylated, moiety of the molecule.
Willey
K P
KP
Institute for Hormone and Fertility Research, Hamburg, Federal Republic of Germany.
Leidenberger
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000378:metabolism* / Q000494:pharmacology; D002918:Chymotrypsin; D003338:Corpus Luteum / Q000378:metabolism; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D004220:Disulfides / Q000032:analysis; D005260:Female; D006639:Histidine; D006801:Humans; D007700:Kinetics; D007985:Leydig Cells / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010958:Plasminogen; D011487:Protein Conformation; D051381:Rats; D011967:Receptors, Gonadotropin / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D013256:Steroids / Q000096:biosynthesis
D006063:Chorionic Gonadotropin; D004220:Disulfides; D011967:Receptors, Gonadotropin; D013256:Steroids; D006639:Histidine; D010958:Plasminogen; D002918:Chymotrypsin; D004047:Diethyl Pyrocarbonate