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Conversion of a gonadotropin-releasing hormone antagonist to an agonist

Nature volume 296, pages 653–655 (1982)Cite this article

Abstract

Gonadotropin-releasing hormone (pyroGlu1-His2-Trp3-Ser4-Tyr5-Gly6-Leu7-Arg8-Pro9-Gly10 amide, GnRH) stimulates pituitary luteinizing hormone (LH) release. An antagonist, D-pyroGlu1-D-Phe2-D-Trp3-D-Lys6-GnRH (GnRH-Ant), binds to the pituitary GnRH receptor and inhibits GnRH-stimulated (10−9 M) gonadotropin release from pituitary cultures (IC50 = 2 × 10×7 M). GnRH-Ant has no measurable agonist activity at concentrations up to 10−6 M. The presence of the D-Lys6 both affords protection against proteolysis and includes an amino group which may be used for derivatization without loss of receptor-binding activity. Formation of the GnRH-Ant dimer by cross-linking of the (lysyl) amino groups of two molecules with ethylene glycol bis(succinimidyl succinate) (EGS) results in a GnRH-Ant dimer joined by a 12–15 Å chain. As the amino terminus on GnRH-Ant is blocked, leaving the D-Lys6 amino group the only reactive group1, the reaction of EGS with GnRH-Ant does not lead to larger polymers. Like the parent compound, this dimer is purely an antagonist. We now show, however, that when antibody (AB) to D-Lys6-GnRH (which cross-reacts with GnRH-Ant) is incubated with excess dimer, a product is formed which consists of a divalent antibody with a GnRH-Ant dimer attached to each arm: AB–((GnRH-Ant)–EGS–(GnRH-Ant))2. In contrast to the parent compounds, this conjugate is an agonist, stimulating LH release from pituitary cultures. Our results suggest that a pure antagonist becomes an agonist when it is capable of bringing two receptor molecules within a critical distance, d (15 Å<d<150 Å). The data indicate that formation of the receptor microaggregate itself is sufficient to stimulate a transmembrane response system.

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Authors and Affiliations

  1. Department of Pharmacology, Duke University Medical Center, Durham, North Carolina, 27710, USA

    P. Michael Conn, Deloris C. Rogers & Timothy Sheffield

  2. Department of Medicine, Duke University Medical Center, Durham, North Carolina, 27710, USA

    James Niedel

  3. Department of Biochemistry, School of Medicine, University of Colorado, 4200 East Ninth Avenue, Denver, Colorado, 80220, USA

    John M. Stewart

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  1. P. Michael Conn
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  2. Deloris C. Rogers
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  4. James Niedel
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  5. Timothy Sheffield
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Conn, P., Rogers, D., Stewart, J. et al. Conversion of a gonadotropin-releasing hormone antagonist to an agonist. Nature 296, 653–655 (1982). https://doi.org/10.1038/296653a0

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