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Exp Clin Endocrinol Diabetes 2004; 112(10): 574-579
DOI: 10.1055/s-2004-830409
Article

J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

The Carboxyterminal Peptide of Chorionic Gonadotropin Facilitates Activation of the Marmoset LH Receptor

T. Müller1 , J. Gromoll1 , A. P. Simula2 , R. Norman2 , R. Sandhowe-Klaverkamp1 , M. Simoni1
  • 1Institute of Reproductive Medicine of the University of Münster, Münster, Germany
  • 2Reproductive Medicine Unit, Department of Obstetrics and Gynaecology, University of Adelaide, The Queen Elizabeth Hospital, Adelaide, Australia
Further Information

Publication History

Received: October 30, 2003 First decision: February 12, 2004

Accepted: March 25, 2004

Publication Date:
02 December 2004 (online)

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Abstract

Luteinizing hormone (LH) and chorionic gonadotropin (CG) are heterodimeric glycoprotein hormones acting on the luteinizing hormone receptor (LHR). In the LHR, which is genomically encoded by eleven exons, exon 10 encodes for the hinge region and its elimination impairs LH action, while CG maintains normal activity. The two gonadotropins differ in the carboxyterminal peptide (CTP) present in CG but absent in LH. Since the marmoset monkey (Callithrix jacchus) LHR naturally lacks exon 10 (LHR type II), we generated two recombinant marmoset gonadotropin preparations, one consisting of the wild type CG and one of truncated CG lacking the CTP (CG-CTP). After calibration in a mouse Leydig cell bioassay against the WHO LH80/522 standard, the ED50 of the CG preparation on a COS7 cell line permanently expressing the marmoset LHR was 4.25 ± 0.21 IU/L (n = 3). Stimulation of the COS7 cell line with equipotent concentrations of CG and CG-CTP resulted in significantly different formation of cAMP (two-way ANOVA, p < 0.001). In particular, cAMP production stimulated by CG-CTP was 3 - 4 times lower compared to CG at the saturating CG concentration (8 IU/L). We conclude, supplementing one current model of LHR activation, that exon 10 might play a permissive role in releasing the constraint of the receptor upon hormone binding, resulting in receptor activation. We speculate that, when exon 10 is lacking, the CTP can overcome its absence and facilitates the opening of the receptor, resulting in normal activation.

Key words

Marmoset monkey - chorionic gonadotropin - LHR - exon 10 - carboxyterminal peptide

References

  • 1 Amato F, Simula A P, Gameau L J, Norman R J. Expression, characterization and immunoassay of recombinant marmoset chorionic gonadotropin dimer and β-subunit.  J Endocrinol. 1998;  159 141-151
    CrossrefPubMedGoogle Scholar
  • 2 Ascoli M, Fanelli F, Segaloff D L. The lutropin/choriogonadotropin receptor, A 2002 Perspective.  Endocr Rev. 2002;  23 141-174
    CrossrefPubMedGoogle Scholar
  • 3 Albanese C, Colin I M, Crowley W F, Ito M, Pestell R G, Weiss J, Jameson J L. The gonadotropin genes: evolution of distinct mechanisms for hormonal control.  Recent Prog Horm Res. 1996;  51 23-58
    PubMedGoogle Scholar
  • 4 Atger M, Misrahi M, Sar S, Le Flem L, Dessen P, Milgrom E. Structure of the human luteinizing hormone-choriogonadotropin receptor gene: unusual promoter and 5′ non-coding regions.  Mol Cell Endocrinol. 1995;  111 113-123
    CrossrefPubMedGoogle Scholar
  • 5 Bahl O P. Human chorionic gonadotropin, its receptor and mechanism of action.  Fed Proc. 1977;  36 2119-2127
    PubMedGoogle Scholar
  • 6 Bousfield G R, Butnev V Y, Gotschall R R, Baker V L, Moore W T. Structural features of mammalian gonadotropins.  Mol Cell Endocrinol. 1996;  125 3-19
    CrossrefPubMedGoogle Scholar
  • 7 Chen H C, Shimohigashi Y, Dufau M L, Catt K J. Characterization and biological properties of chemically deglycosylated human chorionic gonadotropin.  J Biol Chem. 1982;  257 14446-14452
    PubMedGoogle Scholar
  • 8 Dufau M L. The luteinizing hormone receptor.  Ann Rev Physiol. 1998;  60 461-496
    CrossrefPubMedGoogle Scholar
  • 9 Fiddes J C, Goodman H M. The cDNA for the beta-subunit of human chorionic gonadotropin suggests evolution of a gene by read-through into the 3′-untranslated region.  Nature. 1980;  286 684-687
    CrossrefPubMedGoogle Scholar
  • 10 Galet C, Chopineau M, Martinat N, Combarnous Y, Guillou F. Expression of an in vitro biologically active equine LH/CG without C-terminal peptide (CTP) and/or β26 - 110 disulphide bridge.  J Endocrinol. 2000;  167 117-124
    CrossrefPubMedGoogle Scholar
  • 11 Gharib S D, Wierman M E, Shupnik M A, Chin W W. Molecular biology of the pituitary gonadotropins.  Endocr Rev. 1990;  11 177-199
    PubMedGoogle Scholar
  • 12 Gromoll J, Eiholzer U, Nieschlag E, Simoni M. Male hypogonadism caused by a homozygous deletion of exon 10 of the luteinizing hormone receptor: Differential action of human chorionic gonadotropin and LH.  J Clin Endocrinol Metab. 2000;  85 2281-2286
    PubMedGoogle Scholar
  • 13 Gromoll J, Wistuba J, Terwort N, Godmann M, Müller T, Simoni M. A new subclass of the luteinizing hormone/chorionic gonadotropin receptor lacking exon 10 messenger RNA in the New World Monkey (Platyrrhini) lineage.  Biol Reprod. 2003;  69 75-80
    CrossrefPubMedGoogle Scholar
  • 14 Jia X C, Oikawa M, Bo M, Tanaka T, Ny T, Boime I, Hsueh A J. Expression of human luteinizing hormone (LH) receptor: interaction with LH and chorionic gonadotropin from human but not equine, rat, and ovine species.  Mol Endocrinol. 1991;  5 759-768
    PubMedGoogle Scholar
  • 15 Klein J, Lobel L, Pollak S, Ferin M, Xiao E, Sauer M, Lustbader J W. Pharmacokinetics and pharmacodynamics of single-chain recombinant human follicle-stimulating hormone containing the human chorionic gonadotropin carboxyterminal peptide in the rhesus monkey.  Fertil Steril. 2002;  77 1248-1255
    CrossrefPubMedGoogle Scholar
  • 16 Lapthorn A J, Harris D C, Littlejohn A, Lustbader J W, Canfield R E, Machin K J, Morgan F J, Isaacs N W. Crystal structure of human chorionic gonadotropin.  Nature. 1994;  369 455-461
    CrossrefPubMedGoogle Scholar
  • 17 Maston G A, Ruvolo M. Chorionic gonadotropin has a recent origin within primates and an evolutionary history of selection.  Mol Biol Evol. 2002;  19 320-335
    PubMedGoogle Scholar
  • 18 Matzuk C, Boime I. Site-specific mutagenesis defines the intracellular role of the asparagine-linked oligosaccharides of chorionic gonadotropin beta subunit.  J Biol Chem. 1988 a;  263 17106-17111
    PubMedGoogle Scholar
  • 19 Matzuk C, Boime I. The role of the asparagine-linked oligosaccharides of the alpha subunit in the secretion and assembly of human chorionic gonadotropin.  J Cell Biol. 1988 b;  106 1049-1059
    CrossrefPubMedGoogle Scholar
  • 20 Matzuk C, Keene J L, Boime I. Site specificity of the chorionic gonadotropin N-linked oligosaccharides in signal transduction.  J Biol Chem. 1989;  264 2409-2414
    PubMedGoogle Scholar
  • 21 Matzuk M M, Hsueh A JW, Lapolt P, Tsafiri A, Keene J L, Boime I. The biological role of the carboxyterminal extension of human chorionic gonadotropin.  Endocrinology. 1990;  126 376-383
    PubMedGoogle Scholar
  • 22 Merz W E. Biosynthesis of human chorionic gonadotropin: a review.  Eur J Endocrinol. 1996;  135 269-284
    PubMedGoogle Scholar
  • 23 Müller T, Gromoll J, Simoni M. Absence of exon 10 of the human luteinizing hormone (LH) receptor impairs LH, but not human chorionic gonadotropin action.  J Clin Endocrinol Metab. 2003;  88 2242-2249
    CrossrefPubMedGoogle Scholar
  • 24 Müller T, Simoni M, Pekel E, Luetjens C M, Chandolia R, Amato F, Norman R J, Gromoll J. Chorionic gonadotrophin beta subunit mRNA but not luteinising hormone beta subunit mRNA is expressed in the pituitary of the common marmoset (Callithrix jacchus).  J Mol Endocrinol. 2004;  32 115-128
    CrossrefPubMedGoogle Scholar
  • 25 Muyan M, Furuhashi M, Sugahara T, Boime I. The carboxyterminal region of the β-subunits of LH and CG differentially influence secretion and assembly of the heterodimers.  Mol Endocrinol. 1996;  10 1678-1687
    CrossrefPubMedGoogle Scholar
  • 26 Nishi S, Nakabayashi K, Kobilka B, Aaron J W, Hsueh A JW. The ectodomain of the luteinizing hormone receptor interacts with exoloop 2 to constrain the transmembrane region.  J Biol Chem. 2002;  277 3958-3964
    CrossrefPubMedGoogle Scholar
  • 27 Pierce J G, Parsons T F. Glycoprotein hormones: structure and function.  Annual Rev Biochem. 1981;  50 465-495
    CrossrefPubMedGoogle Scholar
  • 28 Ryan R J, Keutmann H T, Charlesworth M C, McCormick D J, Milius R P, Calvo F O, Vutyavanich T. Structure-function relationships of gonadotropins.  Recent Prog Horm Res. 1987;  43 383-429
    PubMedGoogle Scholar
  • 29 Schmidt A, Gromoll J, Weinbauer G F, Galla H-J, Chappel S, Simoni M. Cloning and expression of cynomolgus monkey (Macaca fascicularis) gonadotropins luteinizing hormone and follicle-stimulating hormone and identification of two polymorphic sites in the luteinizing hormone β subunit.  Mol Cell Endocrinol. 1999;  156 73-83
    CrossrefPubMedGoogle Scholar
  • 30 Shao K, Balasubramanian S V, Pope C M, Bahl O P. Effect of individual N-glycosyl chains in the β-subunit on the conformation of human choriogonadotropin.  Mol Cell Endocrinol. 1998;  146 39-48
    CrossrefPubMedGoogle Scholar
  • 31 Simula A P, Amato F, Faast R, Lopata A, Berka J, Norman R J. Luteinizing hormone/chorionic gonadotropin bioactivity in the common marmoset (Callithrix jacchus) is due to a chorionic gonadotropin molecule with a structure intermediate between human chorionic gonadotropin and human luteinizing hormone.  Biol Reprod. 1995;  53 380-389
    PubMedGoogle Scholar
  • 32 Talmadge K, Vamvakopoulos N C, Fiddes J C. Evolution of the genes for the beta subunits of human chorionic gonadotropin and luteinizing hormone.  Nature. 1984;  307 37-40
    CrossrefPubMedGoogle Scholar
  • 33 Themmen A PN, Huhtaniemi I T. Mutations of gonadotropins and gonadotropin receptors: elucidating the physiology and pathophysiology of pituitary-gonadal function.  Endocr Rev. 2000;  21 551-583
    CrossrefPubMedGoogle Scholar
  • 34 Tullner W W. Comparative aspects of primate gonadotropins.  Contrib Primatol. 1974;  3 235-257
    PubMedGoogle Scholar
  • 35 Wickings E J, Qazi M H, Nieschlag E. Determination of biologically active LH in the serum of male rheusus monkeys (Macaca mulatta).  J Reprod Fertil. 1979;  57 497-504
    PubMedGoogle Scholar
  • 36 Wilson C A, Leigh A J, Chapman A J. Gonadotropin glycosylation and function.  J Endocrinol. 1990;  125 3-14
    PubMedGoogle Scholar
  • 37 Zhang F P, Kero J, Huhtaniemi I. The unique exon 10 of the human luteinizing hormone receptor is necessary for expression of the receptor protein at the plasma membrane in the human luteinizing hormone receptor, but deleterious when inserted into the human follicle-stimulating hormone receptor.  Mol Cell Endocrinol. 1998;  142 165-174
    CrossrefPubMedGoogle Scholar
  • 38 Zheng H, Phang T, Yong S S, Ji I, Ji T H. The role of the hinge region of the luteinizing hormone receptor in hormone interaction and signal transduction.  J Biol Chem. 2001;  276 3451-3458
    CrossrefPubMedGoogle Scholar

Prof. Dr. Manuela Simoni

Institute of Reproductive Medicine of the University of Münster

Domagkstraße 11

48129 Münster

Phone: + 492518356444

Fax: + 49 25 18 35 60 93

Email: simoni@uni-muenster.de

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