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Molecular identification and analysis of a novel human corticotropin-releasing factor (CRF) receptor: the CRF2gamma
W A Kostich1, A Chen, K Sperle
1DuPont Merck Research Laboratories, CNS Diseases Research, Wilmington, Delaware 19880, USA.
Abstract:
We report the discovery of a new CRF2 receptor splice isoform found in human brain, which we have termed the CRF2gamma receptor. The CRF2gamma cDNA encodes for a 397-amino acid receptor that has an amino terminus with no significant homology to the already reported alpha- and beta-termini. When expressed in 293-EBNA (Epstein-Barr nuclear antigen) cells, the CRF2gamma receptor responds in a dose-dependent manner to CRF and related peptides with a rank order of potency of urocortin > or = sauvagine>urotensin>r/h CRF, with EC50 values more similar to CRF2alpha than CRF2beta. Equilibrium saturation isotherm analysis with radiolabeled sauvagine reveals a two site/state model for binding to CRF2gamma with a 60 pM Kd high-affinity site and a 5 nM Kd low-affinity site. Analysis of CRF2gamma RNA expression in human brain demonstrates expression in septum and hippocampus, with weaker but detectable expression in amygdala, nucleus accumbens, midbrain, and frontal cortex.