Analyses of gating thermodynamics and effects of deletions in the mechanosensitive channel TREK-1: Comparisons with structural models

Grigory Maksaev, Adina Milac, Andriy Anishkin, H. Robert Guy, Sergei Sukharev

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


TREK-1, a mechanosensitive K channel from the two-pore family (K 2P), is involved in protective regulation of the resting potential in CNS neurons and other tissues. The structure of TREK-1 and the basis of its sensitivity to stretch and variety of lipid-soluble factors remain unknown. Using existing K channel structures as modeling templates, TREK-1 was envisioned as a 2-fold symmetrical complex with the gate formed primarily by the centrally positioned TM2b helices of the second homologous repeat. Opening was modeled as a conical expansion of the barrel separating TM2b's accompanied by extension of TM2a helices with the cytoplasmic TM2a-TM1b connector. Seeking first experimental support to the models we have accomplished thermodynamic analysis of mouse TREK-1 gating and functional testing of several deletion mutants. The predicted increase of the channel in-plane area (ΔA) of ∼5 nm 2 in models was supported by the experimental ΔA of ∼4 nm2 derived from the slope of open probability versus membrane tension in HE K-293T cells and their cytoskeleton-depleted blebs. In response to steps of suction, wild-type channel produced transient currents in cell-attached patches and mostly sustained currents upon patch excision. TREK-1 motifs not present in canonical K channels include divergent cytoplasmic N- and C-termini, and a characteristic 50-residue extracellular loop in the first homologous repeat. Deletion of the extracellular loop (Δ76-124) reduced the average current density in patches, increased spontaneous activity and generated a larger sub-population of high-conductance channels, while activation by tension augmented by arachidonic acid was fully retained. Further deletion of the C-terminal end (Δ76-124/Δ334-411) removed voltage dependency but otherwise produced no additional effect. In an attempt to generate a cysteine-free version of the channel, we mutated two remaining cysteines 159 and 219 in the transmembrane region. C219A did not compromise channel activity, whereas the C159A/S mutants were essentially inactive. Treatment with β-mercaptoethanol suggested that none of these cysteines form functionally-important disulfides.

Original languageEnglish
Pages (from-to)34-42
Number of pages9
Issue number1
StatePublished - 2011


  • Channel domains
  • Gating transition
  • Membrane tension
  • Protein expansion


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