Identification of blocker binding site in mouse TRESK by molecular modeling and mutational studies

Abstract

TWIK (tandem-pore domain weak inward rectifying K+)-related spinal cord K+ channel, TRESK, a member of the tandem-pore domain K+ channel family, is the most recently cloned K-2P channel. TRESK is highly expressed in dorsal root ganglion neuron, a pain sensing neuron, which is a target for analgesics. In this study, a reliable 3D structure for transmembrane (TM) region of mouse TRESK (mTRESK) was constructed, and then the reasonable blocker binding mode of the protein was investigated. The 3D structure of the mTRESK built by homology modeling method was validated with recommend value of stereochemical quality. Based on the validated structure, K+ channel blocker-bound conformation was obtained by molecular docking and 5 ns MD simulation with DPPC lipid bilayer. Our docking study provides the plausible binding mode of known blockers with key interacting residues, especially, F156 and F364. Finally, these modeling results were verified by experimental study with mutation from phenylalanine to alanine (F156A, F364A and F156A/F364A) at the TM2 and TM4. This is the first modeling study for TRESK that can provide structural information of the protein including ligand binding information. These results can be useful in structure based drug design for finding new blockers of the TRESK as potential therapeutic target of pain treatment. (C) 2012 Elsevier B.V. All rights reserved.