Ligand-gated ion channels (LGICs) are natural biosensors generating electrical signals in response to the binding of specific ligands. Creating de novo LGICs for biosensing applications is technically challenging. We have previously designed modified LGICs by linking G protein-coupled receptors (GPCRs) to the Kir6.2 channel. In this article, we extrapolate these design concepts to other channels with different structures and oligomeric states, namely a tetrameric viral Kcv channel and the dimeric mouse TREK-1 channel. After precise engineering of the linker regions, the two ion channels were successfully regulated by a GPCR fused to their N-terminal domain. Two-electrode voltage-clamp recordings showed that Kcv and mTREK-1 fusions were inhibited and activated by GPCR agonists, respectively, and antagonists abolished both effects. Thus, dissimilar ion channels can be allosterically regulated through their N-terminal domains, suggesting that this is a generalizable approach for ion channel engineering.

Distinct classes of potassium channels fused to GPCRs as electrical signaling biosensors / M.D. Garcia-Fernandez, F.C. Chatelain, H. Nury, A. Moroni, C.J. Moreau. - In: CELL REPORTS. METHODS. - ISSN 2667-2375. - 1:8(2021 Dec), pp. 100119.1-100119.15, e1-e3. [10.1016/j.crmeth.2021.100119]

Distinct classes of potassium channels fused to GPCRs as electrical signaling biosensors

A. Moroni
Penultimo
Conceptualization
;
2021

Abstract

Ligand-gated ion channels (LGICs) are natural biosensors generating electrical signals in response to the binding of specific ligands. Creating de novo LGICs for biosensing applications is technically challenging. We have previously designed modified LGICs by linking G protein-coupled receptors (GPCRs) to the Kir6.2 channel. In this article, we extrapolate these design concepts to other channels with different structures and oligomeric states, namely a tetrameric viral Kcv channel and the dimeric mouse TREK-1 channel. After precise engineering of the linker regions, the two ion channels were successfully regulated by a GPCR fused to their N-terminal domain. Two-electrode voltage-clamp recordings showed that Kcv and mTREK-1 fusions were inhibited and activated by GPCR agonists, respectively, and antagonists abolished both effects. Thus, dissimilar ion channels can be allosterically regulated through their N-terminal domains, suggesting that this is a generalizable approach for ion channel engineering.
man-made ligand-gated ion channels; biosensors; protein engineering; electrical signal; ion channels; K2P channels; viral channels; G protein-coupled receptors; allosteric regulation
Settore BIO/09 - Fisiologia
   Bio-inspired electrONIC NOSE interfacing olfactory electrical biosensors and carbon NANOtubes
   NANOZ-ONIC
   European Commission
   Horizon 2020 Framework Programme
   682286
dic-2021
Article (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/1049213
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