Extracellular modulation of TREK-2 activity with nanobodies provides insight into the mechanisms of K2P channel regulation.

Autor:	Rödström KEJ; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK.; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK.; Centre for Medicines Discovery, University of Oxford, Oxford, UK.; Department of Biochemistry, University of Oxford, Oxford, UK., Cloake A; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK., Sörmann J; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK.; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK., Baronina A; Centre for Medicines Discovery, University of Oxford, Oxford, UK., Smith KHM; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK.; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK.; Department of Biochemistry, University of Oxford, Oxford, UK., Pike ACW; Centre for Medicines Discovery, University of Oxford, Oxford, UK., Ang J; Centre for Medicines Discovery, University of Oxford, Oxford, UK., Proks P; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK.; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK., Schewe M; Institute of Physiology, Medical Faculty, Kiel University, Kiel, Germany., Holland-Kaye I; Department of Biochemistry, University of Oxford, Oxford, UK., Bushell SR; Centre for Medicines Discovery, University of Oxford, Oxford, UK., Elliott J; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK., Pardon E; Structural Biology Brussels, Vrije Universiteit Brussel, Brussels, Belgium.; VIB-VUB Center for Structural Biology, VIB, Brussels, Belgium., Baukrowitz T; Institute of Physiology, Medical Faculty, Kiel University, Kiel, Germany., Owens RJ; The Rosalind Franklin Institute, Harwell Campus, Didcot, UK.; Division of Structural Biology, University of Oxford, Oxford, UK., Newstead S; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK.; Department of Biochemistry, University of Oxford, Oxford, UK.; OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford, UK., Steyaert J; Structural Biology Brussels, Vrije Universiteit Brussel, Brussels, Belgium.; VIB-VUB Center for Structural Biology, VIB, Brussels, Belgium., Carpenter EP; Centre for Medicines Discovery, University of Oxford, Oxford, UK. lizcarpen1@gmail.com.; OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford, UK. lizcarpen1@gmail.com., Tucker SJ; Clarendon Laboratory, Department of Physics, University of Oxford, Oxford, UK. stephen.tucker@physics.ox.ac.uk.; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK. stephen.tucker@physics.ox.ac.uk.; OXION Initiative in Ion Channels and Disease, University of Oxford, Oxford, UK. stephen.tucker@physics.ox.ac.uk.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2024 May 16; Vol. 15 (1), pp. 4173. Date of Electronic Publication: 2024 May 16.
DOI:	10.1038/s41467-024-48536-2
Abstrakt:	Potassium channels of the Two-Pore Domain (K2P) subfamily, KCNK1-KCNK18, play crucial roles in controlling the electrical activity of many different cell types and represent attractive therapeutic targets. However, the identification of highly selective small molecule drugs against these channels has been challenging due to the high degree of structural and functional conservation that exists not only between K2P channels, but across the whole K + channel superfamily. To address the issue of selectivity, here we generate camelid antibody fragments (nanobodies) against the TREK-2 (KCNK10) K2P K + channel and identify selective binders including several that directly modulate channel activity. X-ray crystallography and CryoEM data of these nanobodies in complex with TREK-2 also reveal insights into their mechanisms of activation and inhibition via binding to the extracellular loops and Cap domain, as well as their suitability for immunodetection. These structures facilitate design of a biparatropic inhibitory nanobody with markedly improved sensitivity. Together, these results provide important insights into TREK channel gating and provide an alternative, more selective approach to modulation of K2P channel activity via their extracellular domains. (© 2024. The Author(s).)
Databáze:	MEDLINE
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