The ligand-selective domains of corticotropin-releasing factor type 1 and type 2 receptor reside in different extracellular domains: generation of chimeric receptors with a novel ligand-selective profile

J Neurochem. 1999 Aug;73(2):821-9. doi: 10.1046/j.1471-4159.1999.0730821.x.

Abstract

The nonselective human corticotropin-releasing factor (hCRF) receptor 1 (hCRFR1) and the ligand-selective Xenopus CRFR1 (xCRFR1), xCRFR2, and hCRFR2alpha were compared. To understand the interactions of hCRF, ovine CRF (oCRF), rat urocortin (rUcn), and sauvagine, ligands with different affinities for type 1 and type 2 CRFRs, chimeric and mutant receptors of hCRFR1, xCRFR1, hCRFR2alpha, and xCRFR2 were constructed. In cyclic AMP stimulation and CRF-binding assays, it was established that different extracellular regions of CRFR1 and CRFR2 conferred their ligand selectivities. The ligand selectivity of xCRFR1 resided in five N-terminal amino acids, whereas the N-terminus of both CRFR2 proteins did not contribute to their ligand selectivities. Chimeric receptors in which the first extracellular domain of hCRFR1 replaced that of hCRFR2alpha or xCRFR2 showed a similar pharmacological profile to the two parental CRFR2 molecules. Chimeric receptors carrying the N-terminal domain of xCRFR1 linked to hCRFR2alpha or xCRFR2 displayed a novel pharmacological profile. hCRF, rUcn, and sauvagine were bound with high affinity, whereas oCRF was bound with low affinity. Furthermore, when three or five residues of xCRFR1 (Gln76, Gly81, Val83, His88, Leu89; or Gln76, Gly81, Val83) were introduced into receptor chimeras carrying the N-terminus of hCRFR1 linked to xCRFR2, the same novel pharmacology was observed. These data indicate a compensation mechanism of two differentially selecting regions located in different domains of both xCRFR1 and CRFR2.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amphibian Proteins
  • Animals
  • Binding, Competitive / physiology
  • Cells, Cultured
  • Corticotropin-Releasing Hormone / metabolism
  • Corticotropin-Releasing Hormone / pharmacology
  • Cyclic AMP / metabolism
  • Extracellular Space / chemistry
  • Humans
  • Iodine Radioisotopes
  • Kidney / cytology
  • Ligands
  • Molecular Sequence Data
  • Mutagenesis
  • Peptide Hormones
  • Peptides / metabolism
  • Peptides / pharmacology
  • Protein Structure, Tertiary
  • Radioligand Assay
  • Receptors, Corticotropin-Releasing Hormone / chemistry*
  • Receptors, Corticotropin-Releasing Hormone / genetics*
  • Receptors, Corticotropin-Releasing Hormone / metabolism
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Transfection
  • Urocortins
  • Vasodilator Agents / metabolism
  • Vasodilator Agents / pharmacology
  • Xenopus

Substances

  • Amphibian Proteins
  • CRF receptor type 2
  • Iodine Radioisotopes
  • Ligands
  • Peptide Hormones
  • Peptides
  • Receptors, Corticotropin-Releasing Hormone
  • Recombinant Fusion Proteins
  • Urocortins
  • Vasodilator Agents
  • CRF receptor type 1
  • sauvagine
  • Corticotropin-Releasing Hormone
  • Cyclic AMP