NMR structure and dynamics of the agonist dynorphin peptide bound to the human kappa opioid receptor
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. ¹H and15N chemical shift variations indicated that free and bound peptide is in fast exchange in solutions containing 1 mM dynorphin and 0.01...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 112; no. 38; pp. 11852 - 11857 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
National Academy of Sciences
22.09.2015
National Acad Sciences |
Subjects | |
Online Access | Get full text |
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Summary: | 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. ¹H and15N 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 aK
dof ∼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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Author contributions: R.C.S., K.W., and A.M. designed research; C.O., K.L.W., T.D., G.C., and A.M. performed research; C.O., K.L.W., N.D., T.D., B.L.R., and G.C. contributed new reagents/analytic tools; C.O., G.C., R.C.S., K.W., and A.M. analyzed data; and C.O., T.D., B.L.R., G.C., R.C.S., K.W., and A.M. wrote the paper. Edited by Michael F. Summers, Howard Hughes Medical Institute, University of Maryland, Baltimore County, Baltimore, MD, and accepted by the Editorial Board August 13, 2015 (received for review June 2, 2015) |
ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.1510117112 |