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TitleProlyl isomerization controls activation kinetics of a cyclic nucleotide-gated ion channel.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 6401, Year 2020
Publish dateDec 16, 2020
AuthorsPhilipp A M Schmidpeter / Jan Rheinberger / Crina M Nimigean /
PubMed AbstractSthK, a cyclic nucleotide-modulated ion channel from Spirochaeta thermophila, activates slowly upon cAMP increase. This is reminiscent of the slow, cAMP-induced activation reported for the ...SthK, a cyclic nucleotide-modulated ion channel from Spirochaeta thermophila, activates slowly upon cAMP increase. This is reminiscent of the slow, cAMP-induced activation reported for the hyperpolarization-activated and cyclic nucleotide-gated channel HCN2 in the family of so-called pacemaker channels. Here, we investigate slow cAMP-induced activation in purified SthK channels using stopped-flow assays, mutagenesis, enzymatic catalysis and inhibition assays revealing that the cis/trans conformation of a conserved proline in the cyclic nucleotide-binding domain determines the activation kinetics of SthK. We propose that SthK exists in two forms: trans Pro300 SthK with high ligand binding affinity and fast activation, and cis Pro300 SthK with low affinity and slow activation. Following channel activation, the cis/trans equilibrium, catalyzed by prolyl isomerases, is shifted towards trans, while steady-state channel activity is unaffected. Our results reveal prolyl isomerization as a regulatory mechanism for SthK, and potentially eukaryotic HCN channels. This mechanism could contribute to electrical rhythmicity in cells.
External linksNat Commun / PubMed:33328472 / PubMed Central
MethodsEM (single particle)
Resolution3.42 - 6.68 Å
Structure data

21453

6vxz

EMDB-21453, PDB-6vxz:
SthK P300A cyclic nucleotide-gated potassium channel in the closed state, in complex with cAMP
Method: EM (single particle) / Resolution: 3.42 Å

21454

6vy0

EMDB-21454, PDB-6vy0:
SthK P300A cyclic nucleotide-gated potassium channel in a putative active state, in complex with cAMP
Method: EM (single particle) / Resolution: 6.68 Å

Chemicals

ChemComp-CMP:
ADENOSINE-3',5'-CYCLIC-MONOPHOSPHATE

ChemComp-PGW:
(1R)-2-{[(S)-{[(2S)-2,3-dihydroxypropyl]oxy}(hydroxy)phosphoryl]oxy}-1-[(hexadecanoyloxy)methyl]ethyl / phospholipid*YM

Source
  • spirochaeta thermophila (strain atcc 700085 / dsm 6578 / z-1203) (bacteria)
  • spirochaeta thermophila dsm 6578 (bacteria)
KeywordsTRANSPORT PROTEIN

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