http://dx.doi.org/10.25838/d5p-27
Title: | Functional and structural characterization of interactions between opposite subunits in HCN pacemaker channels |
Authors: | Kondapuram, Mahesh Frieg, Benedikt Yüksel, Sezin Schwabe, Tina Sattler, Christian Lelle, Marco Schweinitz, Andrea Schmauder, Ralf Benndorf, Klaus Gohlke, Holger Kusch, Jana |
Keywords: | HCN channel molecular dynamics simulations subunit interaction patch-clamp technique confocal patch-clamp fluorometry |
Issue Date: | 5-Aug-2021 |
Publisher: | N/A |
Abstract: | Hyperpolarization-activated and cyclic nucleotide (HCN) modulated channels are tetrameric cation channels. In each of the four subunits, the intracellular cyclic nucleotide-binding domain (CNBD) is coupled to the transmembrane domain via a helical structure, the C-linker. High-resolution channel structures suggest that the C-linker enables functionally relevant interactions with the opposite subunit, which might be critical for coupling the conformational changes in the CNBD to the channel pore. We combined mutagenesis, patch-clamp technique, confocal patch-clamp fluorometry, and molecular dynamics simulations to show that residue K464 of the C-linker is essential for stabilizing the closed state of the mHCN2 channel by forming interactions with the opposite subunit. MD simulations revealed that both cAMP and K464E induce a rotation of the intracellular domain relative to the channel pore, weakening the autoinhibitory effect of the unoccupied CL-CNBD region. The adopted poses are in excellent agreement with structural results. |
URI: | https://researchdata.hhu.de/handle/entry/98 http://dx.doi.org/10.25838/d5p-27 |
Appears in Collections: | Computational Pharmaceutical Chemistry and Molecular Informatics Group |
File | Description | Size | Format | |
---|---|---|---|---|
DATA_SOURCE_MDsimulation_Modelling.xlsx | MD simulations data | 34.15 kB | Microsoft Excel XML | View/Open |
Data Source Electrophysiology.xlsx | Electrophysiological data | 35.21 kB | Microsoft Excel XML | View/Open |
Data Source Patch-Clamp-Fluorometry.xlsx | Fluorescence data | 9.29 kB | Microsoft Excel XML | View/Open |
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