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000907779 1001_ $$00000-0002-9287-7189$$aKondapuram, Mahesh$$b0
000907779 245__ $$aFunctional and structural characterization of interactions between opposite subunits in HCN pacemaker channels
000907779 260__ $$aLondon$$bSpringer Nature$$c2022
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000907779 520__ $$aHyperpolarization-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.
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000907779 7001_ $$0P:(DE-Juel1)172887$$aFrieg, Benedikt$$b1$$ufzj
000907779 7001_ $$0P:(DE-HGF)0$$aYüksel, Sezin$$b2
000907779 7001_ $$0P:(DE-HGF)0$$aSchwabe, Tina$$b3
000907779 7001_ $$00000-0003-0731-0469$$aSattler, Christian$$b4
000907779 7001_ $$0P:(DE-HGF)0$$aLelle, Marco$$b5
000907779 7001_ $$0P:(DE-HGF)0$$aSchweinitz, Andrea$$b6
000907779 7001_ $$00000-0002-8441-4264$$aSchmauder, Ralf$$b7
000907779 7001_ $$0P:(DE-HGF)0$$aBenndorf, Klaus$$b8
000907779 7001_ $$0P:(DE-Juel1)172663$$aGohlke, Holger$$b9$$eCorresponding author
000907779 7001_ $$00000-0001-7206-8133$$aKusch, Jana$$b10$$eCorresponding author
000907779 773__ $$0PERI:(DE-600)2919698-X$$a10.1038/s42003-022-03360-6$$gVol. 5, no. 1, p. 430$$n1$$p430$$tCommunications biology$$v5$$x2399-3642$$y2022
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