| Structure-based analysis of GPCR function: conformational adaptation of both agonist and receptor upon leukotriene B4 binding to recombinant BLT1. | |
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MedLine Citation:
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PMID: 12787679 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We produced the human leukotriene B(4) (LTB(4)) receptor BLT1, a G-protein-coupled receptor, in Escherichia coli with yields that are sufficient for the first structural characterization of this receptor in solution. Overexpression was achieved through codon optimization and the search for optimal refolding conditions of BLT1 recovered from inclusion bodies. The detergent-solubilized receptor displays a 3D-fold compatible with a seven transmembrane (TM) domain with ca 50% alpha-helix and an essential disulfide bridge (circular dichroism evidence); it binds LTB(4) with K(a)=7.8(+/-0.2)x10(8)M(-1) and a stoichiometric ratio of 0.98(+/-0.02). Antagonistic effects were investigated using a synthetic molecule that shares common structural features with LTB(4). We report evidence that both partners, LTB(4) and BLT1, undergo a rearrangement of their respective conformations upon complex formation: (i) a departure from planarity of the LTB(4) conjugated triene moiety; (ii) a change in the environment of Trp234 (TM-VI helix) and in the exposure of the cytoplasmic region of this transmembrane helix. |
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Authors:
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Jean-Louis Baneres; Aimée Martin; Pierre Hullot; Jean-Pierre Girard; Jean-Claude Rossi; Joseph Parello |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of molecular biology Volume: 329 ISSN: 0022-2836 ISO Abbreviation: J. Mol. Biol. Publication Date: 2003 Jun |
Date Detail:
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Created Date: 2003-06-05 Completed Date: 2003-07-24 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 2985088R Medline TA: J Mol Biol Country: England |
Other Details:
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Languages: eng Pagination: 801-14 Citation Subset: IM |
Affiliation:
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UMR 5074 CNRS, Chimie Biomoléculaire et Interactions Biologiques, Faculté de Pharmacie, 15 Avenue Ch. Flahault, BP 14491, 34093, Cedex 05, Montpellier, France. baneres@pharma.univ-montp1.fr |
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| MeSH Terms | |
Descriptor/Qualifier:
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Circular Dichroism Cloning, Molecular Escherichia coli / metabolism Fluorescence Humans Leukotriene B4 / metabolism* Ligands Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Conformation Receptors, Leukotriene B4 / agonists, chemistry*, genetics, metabolism* Recombinant Proteins / chemistry, metabolism Structure-Activity Relationship Tryptophan / chemistry |
| Chemical | |
Reg. No./Substance:
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0/Ligands; 0/Receptors, Leukotriene B4; 0/Recombinant Proteins; 71160-24-2/Leukotriene B4; 73-22-3/Tryptophan |
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