Functional and structural characterization of interactions between opposite subunits in HCN pacemaker channels

• Main Authors: Benndorf, K. (Author), Frieg, B. (Author), Gohlke, H. (Author), Kondapuram, M. (Author), Kusch, J. (Author), Lelle, M. (Author), Sattler, C. (Author), Schmauder, R. (Author), Schwabe, T. (Author), Schweinitz, A. (Author), Yüksel, S. (Author)
• Format: Article
• Language: English
• Published: NLM (Medline) 2022
• Online Access: View Fulltext in Publisher

 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 (MD) simulations to show that residue K464 of the C-linker is relevant for stabilizing the closed state of the mHCN2 channel by forming interactions with the opposite subunit. MD simulations revealed that in the K464E channel, a rotation of the intracellular domain relative to the channel pore is induced, which is similar to the cAMP-induced rotation, weakening the autoinhibitory effect of the unoccupied CL-CNBD region. We suggest that this CL-CNBD rotation is considerably involved in activation-induced affinity increase but only indirectly involved in gate modulation. The adopted poses shown herein are in excellent agreement with previous structural results. © 2022. The Author(s).