K2P channel gating mechanisms revealed by structures of TREK-2 and a complex with Prozac

Yin Yao Dong; Ashley C W Pike; Alexandra Mackenzie; Conor McClenaghan; Prafulla Aryal; Liang Dong; Andrew Quigley; Mariana Grieben; Solenne Goubin; Shubhashish Mukhopadhyay; Gian Filippo Ruda; Michael V Clausen; Lishuang Cao; Paul E Brennan; Nicola A Burgess-Brown; Mark S P Sansom; Stephen J Tucker; Elisabeth P Carpenter

doi:10.1126/science.1261512

K2P channel gating mechanisms revealed by structures of TREK-2 and a complex with Prozac

Science. 2015 Mar 13;347(6227):1256-9. doi: 10.1126/science.1261512.

Authors

Yin Yao Dong¹, Ashley C W Pike¹, Alexandra Mackenzie², Conor McClenaghan³, Prafulla Aryal⁴, Liang Dong¹, Andrew Quigley¹, Mariana Grieben¹, Solenne Goubin¹, Shubhashish Mukhopadhyay¹, Gian Filippo Ruda⁵, Michael V Clausen⁶, Lishuang Cao⁷, Paul E Brennan⁵, Nicola A Burgess-Brown¹, Mark S P Sansom⁸, Stephen J Tucker⁹, Elisabeth P Carpenter¹⁰

Affiliations

¹ Structural Genomics Consortium, University of Oxford, Oxford OX3 7DQ, UK.
² Structural Genomics Consortium, University of Oxford, Oxford OX3 7DQ, UK. Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, UK.
³ Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, UK. OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford OX1 3PN, UK.
⁴ Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, UK. OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford OX1 3PN, UK. Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK.
⁵ Structural Genomics Consortium, University of Oxford, Oxford OX3 7DQ, UK. Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7FZ, UK.
⁶ Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, UK.
⁷ Pfizer Neusentis, Granta Park, Cambridge CB21 6GS, UK.
⁸ OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford OX1 3PN, UK. Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK.
⁹ Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, UK. OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford OX1 3PN, UK. liz.carpenter@sgc.ox.ac.uk stephen.tucker@physics.ox.ac.uk.
¹⁰ Structural Genomics Consortium, University of Oxford, Oxford OX3 7DQ, UK. OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford OX1 3PN, UK. liz.carpenter@sgc.ox.ac.uk stephen.tucker@physics.ox.ac.uk.

Abstract

TREK-2 (KCNK10/K2P10), a two-pore domain potassium (K2P) channel, is gated by multiple stimuli such as stretch, fatty acids, and pH and by several drugs. However, the mechanisms that control channel gating are unclear. Here we present crystal structures of the human TREK-2 channel (up to 3.4 angstrom resolution) in two conformations and in complex with norfluoxetine, the active metabolite of fluoxetine (Prozac) and a state-dependent blocker of TREK channels. Norfluoxetine binds within intramembrane fenestrations found in only one of these two conformations. Channel activation by arachidonic acid and mechanical stretch involves conversion between these states through movement of the pore-lining helices. These results provide an explanation for TREK channel mechanosensitivity, regulation by diverse stimuli, and possible off-target effects of the serotonin reuptake inhibitor Prozac.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Arachidonic Acid / pharmacology
Binding Sites
Crystallography, X-Ray
Fluoxetine / analogs & derivatives
Fluoxetine / chemistry
Fluoxetine / metabolism
Fluoxetine / pharmacology
Humans
Ion Channel Gating*
Models, Molecular
Molecular Dynamics Simulation
Molecular Sequence Data
Potassium / metabolism
Potassium Channels, Tandem Pore Domain / antagonists & inhibitors
Potassium Channels, Tandem Pore Domain / chemistry*
Potassium Channels, Tandem Pore Domain / metabolism
Protein Conformation
Protein Folding
Protein Structure, Secondary
Protein Structure, Tertiary

Substances

KCNK10 protein, human
Potassium Channels, Tandem Pore Domain
Fluoxetine
Arachidonic Acid
norfluoxetine
Potassium

Associated data

PDB/4BW5
PDB/4XDJ
PDB/4XDK
PDB/4XDL

Abstract

Publication types

MeSH terms

Substances

Associated data

Grants and funding