2o8z
From Proteopedia
(New page: 200px<br /><applet load="2o8z" size="350" color="white" frame="true" align="right" spinBox="true" caption="2o8z" /> '''Bound Structure of CRF1 Extracellular Domain...) |
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==Overview== | ==Overview== | ||
| - | Natural peptide agonists of corticotrophin-releasing factor (CRF) | + | 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. |
==About this Structure== | ==About this Structure== | ||
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NMR structural characterization of a minimal peptide antagonist bound to the extracellular domain of the corticotropin-releasing factor1 receptor., Mesleh MF, Shirley WA, Heise CE, Ling N, Maki RA, Laura RP, J Biol Chem. 2007 Mar 2;282(9):6338-46. Epub 2006 Dec 27. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17192263 17192263] | NMR structural characterization of a minimal peptide antagonist bound to the extracellular domain of the corticotropin-releasing factor1 receptor., Mesleh MF, Shirley WA, Heise CE, Ling N, Maki RA, Laura RP, J Biol Chem. 2007 Mar 2;282(9):6338-46. Epub 2006 Dec 27. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17192263 17192263] | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
| - | [[Category: Heise, C | + | [[Category: Heise, C E.]] |
| - | [[Category: Laura, R | + | [[Category: Laura, R P.]] |
[[Category: Ling, N.]] | [[Category: Ling, N.]] | ||
| - | [[Category: Maki, R | + | [[Category: Maki, R A.]] |
| - | [[Category: Mesleh, M | + | [[Category: Mesleh, M F.]] |
| - | [[Category: Shirley, W | + | [[Category: Shirley, W A.]] |
[[Category: ACE]] | [[Category: ACE]] | ||
[[Category: crf]] | [[Category: crf]] | ||
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[[Category: peptide ligand]] | [[Category: peptide ligand]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:15:49 2008'' |
Revision as of 16:15, 21 February 2008
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Bound Structure of CRF1 Extracellular Domain Antagonist
Overview
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.
About this Structure
2O8Z is a Protein complex structure of sequences from [1] with as ligand. Full crystallographic information is available from OCA.
Reference
NMR structural characterization of a minimal peptide antagonist bound to the extracellular domain of the corticotropin-releasing factor1 receptor., Mesleh MF, Shirley WA, Heise CE, Ling N, Maki RA, Laura RP, J Biol Chem. 2007 Mar 2;282(9):6338-46. Epub 2006 Dec 27. PMID:17192263
Page seeded by OCA on Thu Feb 21 18:15:49 2008
Categories: Protein complex | Heise, C E. | Laura, R P. | Ling, N. | Maki, R A. | Mesleh, M F. | Shirley, W A. | ACE | Crf | Ecd | Extracellular domain | Gpcr | Helical | Peptide ligand
