Structure of the Nanobody-Stabilized Active State of the Kappa Opioid Receptor

Tao Che; Susruta Majumdar; Saheem A Zaidi; Pauline Ondachi; John D McCorvy; Sheng Wang; Philip D Mosier; Rajendra Uprety; Eyal Vardy; Brian E Krumm; Gye Won Han; Ming-Yue Lee; Els Pardon; Jan Steyaert; Xi-Ping Huang; Ryan T Strachan; Alexandra R Tribo; Gavril W Pasternak; F Ivy Carroll; Raymond C Stevens; Vadim Cherezov; Vsevolod Katritch; Daniel Wacker; Bryan L Roth

doi:10.1016/j.cell.2017.12.011

Structure of the Nanobody-Stabilized Active State of the Kappa Opioid Receptor

Cell. 2018 Jan 11;172(1-2):55-67.e15. doi: 10.1016/j.cell.2017.12.011. Epub 2018 Jan 4.

Authors

Tao Che¹, Susruta Majumdar², Saheem A Zaidi³, Pauline Ondachi⁴, John D McCorvy¹, Sheng Wang¹, Philip D Mosier⁵, Rajendra Uprety², Eyal Vardy¹, Brian E Krumm¹, Gye Won Han⁶, Ming-Yue Lee⁷, Els Pardon⁸, Jan Steyaert⁸, Xi-Ping Huang⁹, Ryan T Strachan¹, Alexandra R Tribo¹, Gavril W Pasternak², F Ivy Carroll⁴, Raymond C Stevens¹⁰, Vadim Cherezov⁶, Vsevolod Katritch¹⁰, Daniel Wacker¹¹, Bryan L Roth¹²

Affiliations

¹ Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.
² Molecular Pharmacology Program and Department of Neurology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
³ Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
⁴ Center for Organic and Medicinal Chemistry, Research Triangle Institute, Research Triangle Park, NC 27709, USA.
⁵ Department of Medicinal Chemistry and Institute for Structural Biology, Drug Discovery and Development, Virginia Commonweath University, Richmond, VA 23298, USA.
⁶ Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
⁷ Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA; School of Molecular Sciences, Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA; Institute of Natural Resources and Environmental Audits, Nanjing Audit University, Nanjing, China.
⁸ Structural Biology Brussels, Vrije Universiteit Brussel, 1050 Brussels, Belgium; VIB-VUB Center for Structural Biology, VIB, 1050 Brussels, Belgium.
⁹ National Institute of Mental Health Psychoactive Drug Screening Program (NIMH PDSP), School of Medicine, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.
¹⁰ Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA; Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
¹¹ Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA. Electronic address: dwacker@email.unc.edu.
¹² Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA; National Institute of Mental Health Psychoactive Drug Screening Program (NIMH PDSP), School of Medicine, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA; Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: bryan_roth@med.unc.edu.

Abstract

The κ-opioid receptor (KOP) mediates the actions of opioids with hallucinogenic, dysphoric, and analgesic activities. The design of KOP analgesics devoid of hallucinatory and dysphoric effects has been hindered by an incomplete structural and mechanistic understanding of KOP agonist actions. Here, we provide a crystal structure of human KOP in complex with the potent epoxymorphinan opioid agonist MP1104 and an active-state-stabilizing nanobody. Comparisons between inactive- and active-state opioid receptor structures reveal substantial conformational changes in the binding pocket and intracellular and extracellular regions. Extensive structural analysis and experimental validation illuminate key residues that propagate larger-scale structural rearrangements and transducer binding that, collectively, elucidate the structural determinants of KOP pharmacology, function, and biased signaling. These molecular insights promise to accelerate the structure-guided design of safer and more effective κ-opioid receptor therapeutics.

Keywords: GPCR; active state; addiction; crystallography; morphinan; nanobody; opioid receptor; structure-function.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Analgesics / chemistry
Analgesics / pharmacology
Animals
Binding Sites
HEK293 Cells
Humans
Molecular Docking Simulation*
Molecular Dynamics Simulation
Morphinans / chemistry
Morphinans / pharmacology
Protein Binding
Protein Stability
Receptors, Opioid, kappa / agonists
Receptors, Opioid, kappa / chemistry*
Receptors, Opioid, kappa / metabolism
Sf9 Cells
Spodoptera

Substances

17-cyclopropylmethyl-3-hydroxy-4,5-epoxy-7,8-en-6-((3'-iodo)benzamido)morphinan
Analgesics
Morphinans
Receptors, Opioid, kappa

Abstract

Publication types

MeSH terms

Substances

Grants and funding