Chimaeric gonadotropin-releasing hormone (GnRH) peptides with improved affinity for the catfish (Clarias gariepinus) GnRH receptor.
The gonadotropin-releasing hormone (GnRH) receptor in catfish differs from its mammalian counterparts in showing a very low affinity for the hypothalamic GnRH form [i.e. catfish GnRH (cfGnRH)] and a very high affinity for the highly conserved mesencephalic GnRH, chicken GnRH-II (cGnRH-II). In the present study we investigated the molecular interactions between ligand and receptor involved in determining the ligand selectivity of the catfish GnRH receptor. Studies on the binding characteristics of the catfish GnRH receptor for cfGnRH and cGnRH-II as well as for mammalian GnRH (mGnRH) and synthetic chimaeric GnRHs, differing at positions 5, 7 and 8, revealed that the low affinity of the catfish receptor for cfGnRH can be improved by replacing Leu(7) by a tryptophan residue and/or Asn(8) by either a tyrosine or an arginine residue. Testing cfGnRH and cGnRH-II as well as mGnRH and the chimaeric GnRHs on Asp(304)-->Ala, Asp(304)-->Glu and Asp(304)-->Asn mutant catfish GnRH receptors revealed that Asp(304) of the catfish receptor mediates the recognition of Arg(8) in mGnRH, as well as in the chimaeric peptides [Arg(8)]cfGnRH and [Arg(8)]cGnRH-II, but seems to be less important for the recognition of Tyr(8) in cGnRH-II. On the basis of these results, a three-dimensional model for the binding of [Arg(8)]cGnRH-II to the catfish GnRH receptor is proposed.
Blomenröhr
Marion
M
Department of Experimental Zoology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
ter Laak
Ton
T
Kühne
Ronald
R
Beyermann
Michael
M
Hund
Eveline
E
Bogerd
Jan
J
Leurs
Rob
R
D000409:Alanine / Q000737:chemistry; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D001665:Binding Sites; D002397:Catfishes; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D018698:Glutamic Acid / Q000737:chemistry; D007987:Gonadotropin-Releasing Hormone / Q000737:chemistry*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011966:Receptors, LHRH / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014162:Transfection; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
D018076:DNA, Complementary; D008024:Ligands; D010455:Peptides; D011966:Receptors, LHRH; D011993:Recombinant Fusion Proteins; D001224:Aspartic Acid; D007987:Gonadotropin-Releasing Hormone; D018698:Glutamic Acid; D014443:Tyrosine; D014364:Tryptophan; D000409:Alanine