2O8Z
Bound Structure of CRF1 Extracellular Domain Antagonist
Summary for 2O8Z
| Entry DOI | 10.2210/pdb2o8z/pdb |
| Descriptor | cCRF(30-41) Peptide (1 entity in total) |
| Functional Keywords | helical, crf, peptide ligand, gpcr, ecd, extracellular domain, neuropeptide |
| Total number of polymer chains | 1 |
| Total formula weight | 1509.84 |
| Authors | Mesleh, M.F.,Shirley, W.A.,Heise, C.E.,Ling, N.,Maki, R.A.,Laura, R.P. (deposition date: 2006-12-12, release date: 2006-12-26, Last modification date: 2024-11-13) |
| Primary citation | Mesleh, M.F.,Shirley, W.A.,Heise, C.E.,Ling, N.,Maki, R.A.,Laura, R.P. NMR structural characterization of a minimal peptide antagonist bound to the extracellular domain of the corticotropin-releasing factor1 receptor. J.Biol.Chem., 282:6338-6346, 2007 Cited by PubMed Abstract: Natural peptide agonists of corticotrophin-releasing factor (CRF) receptors bind to the receptor by a two-site mechanism as follows: the carboxyl end of the ligand binds the N-terminal extracellular domain (ECD) of the receptor and the amino portion of the ligand binds the extracellular face of the seven transmembrane region. Recently, peptide antagonists homologous to the 12 C-terminal residues of CRF have been derived, which bind the CRF(1) receptor through an interaction with the ECD. Here we characterized the binding of a minimal 12-residue peptide antagonist while bound to the isolated ECD of the CRF(1) receptor. We have expressed and purified soluble and properly folded ECD independent from the seven-transmembrane region as a thioredoxin fusion protein in Escherichia coli. A model of the peptide antagonist, cyclic corticotrophin-releasing factor residues 30-41 (cCRF(30-41)), was calculated while bound to the recombinant ECD using transferred nuclear Overhauser effect spectroscopy. Although the peptide is unstructured in solution, it adopts an alpha-helical conformation when bound to the ECD. Residues of cCRF(30-41) comprising the binding interface with the ECD were mapped using saturation transfer difference NMR. Two hydrophobic residues (Met(38) and Ile(41)) as well as two amide groups (Asn(34) and the C-terminal amide) on one face of the helix defined the binding epitope of the antagonist. This epitope may be used as a starting point for development of non-peptide antagonists targeting the ECD of this receptor. PubMed: 17192263DOI: 10.1074/jbc.M609816200 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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