Functional and structural characterization of interactions between opposite subunits in HCN pacemaker channels.
Commun Biol
; 5(1): 430, 2022 05 09.
Article
in En
| MEDLINE
| ID: mdl-35534535
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 (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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels
/
Nucleotides, Cyclic
Language:
En
Journal:
Commun Biol
Year:
2022
Document type:
Article
Affiliation country:
Germany