title: Structural requirements for the interaction of sheep insulin-like factor 3 with relaxin receptors in rat atria.
abstract: Relaxin is a peptide with various reproductive and nonreproductive functions. The site for the peptide-receptor interaction contains two arginines (Arg) and an isoleucine (Ile) or valine (Val) residue in the B-chain with a configuration of -Arg-X-X-X-Arg-X-X-Ile/Val-X-. The sheep insulin-like peptide 3 (INSL3), a structural homologue of relaxin, also contains the n, n+4 arginines in the B-chain but they are displaced towards the carboxyl terminus by four residues (-X-X-X-X-Arg-X-X-Val-Arg-). Human INSL3 increases the activity of human relaxin in mouse bioassays. Here, we investigated whether sheep synthetic INSL3 affects the relaxin activity in rat atria. INSL3 lacked relaxin-like agonist activity but blocked the activity of relaxin and competed for relaxin binding sites at high concentrations. We also synthesized analogues of INSL3, with amino acid substitutions in the arginine-binding region. Analogues A, D and E, which have the arginines in positions identical to relaxin, showed weak relaxin-like agonist activity. These results suggest that other sites in the relaxin molecule are involved in high-affinity peptide-receptor interaction for the production of the relaxin biological responses.
authors.affiliation: Department of Pharmacology, Monash University, P.O. Box 13E, Clayton 3800, Victoria, Australia
authors.forename: Yean Yeow
authors.initials: YY
authors.lastname: Tan
authors.forename: Nicola F
authors.initials: NF
authors.lastname: Dawson
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Kompa
authors.forename: Courtney P
authors.initials: CP
authors.lastname: Bond
authors.forename: Antonia
authors.initials: A
authors.lastname: Claasz
authors.forename: John D
authors.initials: JD
authors.lastname: Wade
authors.forename: Geoffrey W
authors.initials: GW
authors.lastname: Tregear
authors.forename: Roger J
authors.initials: RJ
authors.lastname: Summers
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D001667:Binding, Competitive; D018360:Crystallography, X-Ray; D006325:Heart Atria / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide / Q000096:biosynthesis / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012065:Relaxin / Q000737:chemistry / Q000378:metabolism*; D012756:Sheep
chemical_list: D007328:Insulin; C084797:Leydig insulin-like protein; D011506:Proteins; C095068:RLN2 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D011994:Recombinant Proteins; C033395:relaxin receptors; D012065:Relaxin