Al-Moubarak, Ehab, Veale, Emma L., Mathie, Alistair (2019) Pharmacologically Reversible, Loss of Function Mutations in the tm2 and tm4 Inner Pore Helices of Trek-1 k2p Channels. Scientific Reports, 9 . Article Number 12394. ISSN 2045-2322. (doi:10.1038/s41598-019-48855-1) (KAR id:76214)
PDF
Publisher pdf
Language: English
This work is licensed under a Creative Commons Attribution 4.0 International License.
|
|
Download this file (PDF/3MB) |
Preview |
Request a format suitable for use with assistive technology e.g. a screenreader | |
Official URL: https://doi.org/10.1038/s41598-019-48855-1 |
Abstract
A better understanding of the gating of TREK two pore domain potassium (K2P) channels and their activation by compounds such as the negatively charged activator, flufenamic acid (FFA) is critical in the search for more potent and selective activators of these channels. Currents through wild-type and mutated human K2P channels expressed in tsA201 cells were measured using whole-cell patch-clamp recordings in the presence and absence of FFA. Mutation of the TM2.6 residue of TREK-1 to a phenylalanine (G171F) and a similar mutation of TM4.6 (A286F) substantially reduced current through TREK-1 channels. In complementary experiments, replacing the natural F residues at the equivalent position in TRESK channels, significantly enhanced current. Known, gain of function mutations of TREK-1 (G137I, Y284A) recovered current through these mutated channels. This reduction in current could be also be reversed pharmacologically, by FFA. However, an appropriate length MTS (MethaneThioSulfonate) cross-linking reagent (MTS14) restricted the activation of TREK-1_A286C channels by repeated application of FFA. This suggests that the cross-linker stabilises the channel in a conformation which blunts FFA activation. Pharmacologically reversible mutations of TREK channels will help to clarify the importance of these channels in pathophysiological conditions such as pain and depression.
Item Type: | Article |
---|---|
DOI/Identification number: | 10.1038/s41598-019-48855-1 |
Divisions: | Divisions > Division of Natural Sciences > Medway School of Pharmacy |
Depositing User: | Alistair Mathie |
Date Deposited: | 05 Sep 2019 13:45 UTC |
Last Modified: | 04 Mar 2024 18:49 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/76214 (The current URI for this page, for reference purposes) |
- Link to SensusAccess
- Export to:
- RefWorks
- EPrints3 XML
- BibTeX
- CSV
- Depositors only (login required):