The Polysite Pharmacology of TREK K2P Channels

Pope, Lianne; Minor, Daniel L.

doi:10.1007/978-981-16-4254-8_4

Cited by 8 publications

(9 citation statements)

References 130 publications

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“…But unlike other TREK structures where this M4 tryptophan has rotated inwards, here the adjacent residues on PH1 (F164) also flips orientation to interact with the top of M4 to stabilise this displacement (Fig 4F). This demonstrates a mechanism whereby movement of the M4 helix may communicate with PH1 to influence the selectivity filter gate and highlights the importance of this 'K2P modulator pocket' in channel gating (36). 5E-F).…”

Section: Immunodetection Of Trek-2 Expression At the Cell Surfacementioning

confidence: 82%

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

Rödström,

Cloake,

Sörmann

et al. 2023

Preprint

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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, we generated camelid antibody fragments (nanobodies) against the TREK-2 (KCNK10) K2P K+ channel and identified selective binders including several that directly modulate channel activity. Crystal structures 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 tools therefore provide important insights into TREK channel gating and a more selective approach to the modulation of K2P channel activity via their extracellular domains.

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Section: Immunodetection Of Trek-2 Expression At the Cell Surfacementioning

confidence: 82%

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

Rödström,

Cloake,

Sörmann

et al. 2023

Preprint

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“…The polysite nature of K 2P pharmacology raises the possibility for synergistic action among modulators acting on the various effector binding sites 16,57 . However, apart from the observed independence of the K 2P modulator pocket (MP) and Keystone inhibitor site located under the K 2P cap 20 , the degree of coupling among the various modulator binding sites has been largely unexplored.…”

Section: Discussionmentioning

confidence: 99%

Development of covalent chemogenetic K_2Pchannel activators

Deal,

Lee,

Mondal

et al. 2023

Preprint

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K2Ppotassium channels regulate excitability by affecting cellular resting membrane potential in the brain, cardiovascular system, immune cells, and sensory organs. Despite their important roles in anesthesia, arrhythmia, pain, hypertension, sleep, and migraine, the ability to control K2Pfunction remains limited. Here, we describe a chemogenetic strategy termed CATKLAMP (CovalentActivation ofTREK familyK+channels to cLAmpMembranePotential) that leverages the discovery of a site in the K2Pmodulator pocket that reacts with electrophile bearing derivatives of a TREK subfamily small molecule activator, ML335, to activate the channel irreversibly. We show that the CATKLAMP strategy can be used to probe fundamental aspects of K2Pfunction, as a switch to silence neuronal firing, and is applicable to all TREK subfamily members. Together, our findings exemplify a new means to alter K2Pchannel activity that should facilitate studies both molecular and systems level studies of K2Pfunction and enable the search for new K2Pmodulators.

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“…Among the pharmacological agents acting on the TREK channels, a widely used anti-depressant fluoxetine and its active metabolite norfluoxetine (NFx) are known as effective inhibitors on both TREK-1 and -2 [ 46 , 47 ]. Although not the principal targets of the antidepressant, TREK channel inhibition by NFx has provided important insights into the conformational changes associated with the gating mechanism of the TREK channels [ 47 , 48 ]. Therefore, we examined whether the I TREK2 maximally activated by Ech A combined with 2-APB was still inhibited by NFx.…”

Section: Resultsmentioning

confidence: 99%

“…As mentioned above, TREK-2 has “up” and “down” positions of the M4 helix. The “down” state creates the fenestration site just below the second pore helix (P2) that is open to the center of the membrane [ 48 ]. The TREK inhibitor NFx binds with the fenestration site defined by the lower part of the P2 pore helix and the “down” state M4 [ 63 ].…”

Section: Discussionmentioning

confidence: 99%

Multiple Effects of Echinochrome A on Selected Ion Channels Implicated in Skin Physiology

et al. 2023

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Echinochrome A (Ech A), a naphthoquinoid pigment from sea urchins, is known to have anti-inflammatory and analgesic effects that have been suggested to be mediated by antioxidant activity and intracellular signaling modulation. In addition to these mechanisms, the ion channels in keratinocytes, immune cells, and nociceptive neurons may be the target for the pharmacological effects. Here, using the patch clamp technique, we investigated the effects of Ech A on the Ca2+-permeable TRPV3, TRPV1 and Orai1 channels and the two-pore domain K+ (K2P) channels (TREK/TRAAK, TASK-1, and TRESK) overexpressed in HEK 293 cells. Ech A inhibited both the TRPV3 and Orai1 currents, with IC50 levels of 2.1 and 2.4 μM, respectively. The capsaicin-activated TRPV1 current was slightly augmented by Ech A. Ech A alone did not change the amplitude of the TREK-2 current (ITREK2), but pretreatments with Ech A markedly facilitated ITREK2 activation by 2-APB, arachidonic acid (AA), and acidic extracellular pH (pHe). Similar facilitation effects of Ech A on TREK-1 and TRAAK were observed when they were stimulated with 2-APB and AA, respectively. On the contrary, Ech A did not affect the TRESK and TASK-1 currents. Interestingly, the ITREK2 maximally activated by the combined application of 2-APB and Ech A was not inhibited by norfluoxetine but was still completely inhibited by ruthenium red. The selective loss of sensitivity to norfluoxetine suggested an altered molecular conformation of TREK-2 by Ech A. We conclude that the Ech A-induced inhibition of the Ca2+-permeable cation channels and the facilitation of the TREK/TRAAK K2P channels may underlie the analgesic and anti-inflammatory effects of Ech A.

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The Polysite Pharmacology of TREK K2P Channels

Cited by 8 publications

References 130 publications

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

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

Development of covalent chemogenetic K_2Pchannel activators

Multiple Effects of Echinochrome A on Selected Ion Channels Implicated in Skin Physiology

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The Polysite Pharmacology of TREK K2P Channels

Cited by 8 publications

References 130 publications

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

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

Development of covalent chemogenetic K2Pchannel activators

Multiple Effects of Echinochrome A on Selected Ion Channels Implicated in Skin Physiology

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Development of covalent chemogenetic K_2Pchannel activators