title: NMR structure and dynamics of the agonist dynorphin peptide bound to the human kappa opioid receptor.
abstract: The structure of the dynorphin (1-13) peptide (dynorphin) bound to the human kappa opioid receptor (KOR) has been determined by liquid-state NMR spectroscopy. (1)H and (15)N chemical shift variations indicated that free and bound peptide is in fast exchange in solutions containing 1 mM dynorphin and 0.01 mM KOR. Radioligand binding indicated an intermediate-affinity interaction, with a Kd of ∼200 nM. Transferred nuclear Overhauser enhancement spectroscopy was used to determine the structure of bound dynorphin. The N-terminal opioid signature, YGGF, was observed to be flexibly disordered, the central part of the peptide from L5 to R9 to form a helical turn, and the C-terminal segment from P10 to K13 to be flexibly disordered in this intermediate-affinity bound state. Combining molecular modeling with NMR provided an initial framework for understanding multistep activation of a G protein-coupled receptor by its cognate peptide ligand.
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037; Departments of Biological Sciences and Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089;
authors.forename: Casey
authors.initials: C
authors.lastname: O'Connor
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037; Departments of Biological Sciences and Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089; Department of Pharmacology, University of North Carolina Chapel Hill Medical School, Chapel Hill, NC 27514;
authors.forename: Kate L
authors.initials: KL
authors.lastname: White
authors.affiliation: Institute of Pharmacology and Structural Biology, CNRS and Université de Toulouse-Paul Sabatier, 31077 Toulouse, France;
authors.forename: Nathalie
authors.initials: N
authors.lastname: Doncescu
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Tatiana
authors.initials: T
authors.lastname: Didenko
authors.affiliation: Department of Pharmacology, University of North Carolina Chapel Hill Medical School, Chapel Hill, NC 27514;
authors.forename: Bryan L
authors.initials: BL
authors.lastname: Roth
authors.affiliation: Institute of Pharmacology and Structural Biology, CNRS and Université de Toulouse-Paul Sabatier, 31077 Toulouse, France;
authors.forename: Georges
authors.initials: G
authors.lastname: Czaplicki
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037; Departments of Biological Sciences and Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089; stevens@usc.edu wuthrich@scripps.edu alain.milon@ipbs.fr.
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Stevens
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Molecular Biology and Biophysics, ETH Zurich, CH-8093 Zurich, Switzerland; The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037 stevens@usc.edu wuthrich@scripps.edu alain.milon@ipbs.fr.
authors.forename: Kurt
authors.initials: K
authors.lastname: Wüthrich
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037; Institute of Pharmacology and Structural Biology, CNRS and Université de Toulouse-Paul Sabatier, 31077 Toulouse, France; stevens@usc.edu wuthrich@scripps.edu alain.milon@ipbs.fr.
authors.forename: Alain
authors.initials: A
authors.lastname: Milon
mesh_terms: D000595:Amino Acid Sequence; D004399:Dynorphins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D010455:Peptides; D010880:Piperidines / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017473:Receptors, Opioid, kappa / Q000819:agonists* / Q000737:chemistry / Q000378:metabolism*; D044005:Tetrahydroisoquinolines / Q000737:chemistry; D013997:Time Factors
chemical_list: C433814:7-hydroxy-N-(1-((4-(3-hydroxyphenyl)-3,4-dimethyl-1-piperidinyl)methyl)-2-methylpropyl)-1,2,3,4-tetrahydro-3-isoquinolinecarboxamide; D008024:Ligands; D009587:Nitrogen Isotopes; D010446:Peptide Fragments; D010455:Peptides; D010880:Piperidines; D017473:Receptors, Opioid, kappa; D044005:Tetrahydroisoquinolines; C026843:dynorphin (1-13); D004399:Dynorphins
keywords: 15N relaxation; GPCR activation; ligand binding affinity; molecular dynamics simulations; transferred NOE