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PMID: 16407566 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

The Kv2.1 C terminus can autonomously transfer Kv2.1-like phosphorylation-dependent localization, voltage-dependent gating, and muscarinic modulation to diverse Kv channels.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 26 ·No. 2 ·2006-01-11 ·Pages 685-95

Mohapatra DP, Trimmer JS

Abstract

Modulation of K+ channels is widely used to dynamically regulate neuronal membrane excitability. The voltage-gated K+ channel Kv2.1 is an abundant delayed rectifier K+ (IK) channel expressed at high levels in many types of mammalian central neurons where it regulates diverse aspects of membrane excitability. Neuronal Kv2.1 is constitutively phosphorylated, localized in high-density somatodendritic clusters, and has a relatively depolarized voltage dependence of activation. Here, we show that the clustering and voltage-dependent gating of endogenous Kv2.1 in cultured rat hippocampal neurons are modulated by cholinergic stimulation, a common form of neuromodulation. The properties of neuronal Kv2.1 are recapitulated in recombinant Kv2.1 expressed in human embryonic kidney 293 (HEK293) cells, but not COS-1 cells, because of cell background-specific differences in Kv2.1 phosphorylation. As in neurons, Kv2.1 in HEK293 cells is dynamically regulated by cholinergic stimulation, which leads to Ca2+/calcineurin-dependent dephosphorylation of Kv2.1, dispersion of channel clusters, and hyperpolarizing shifts in the voltage-dependent gating properties of the channel. Immunocytochemical, biochemical, and biophysical analyses of chimeric Kv channels show that the Kv2.1 cytoplasmic C-terminal domain can act as an autonomous domain sufficient to transfer Kv2.1-like clustering, voltage-dependent activation, and cholinergic modulation to diverse Kv channels. These findings provide novel mechanistic insights into cholinergic modulation of ion channels and regulation of the localization and voltage-dependent gating properties of the abundant neuronal Kv2.1 channel by cholinergic and other neuromodulatory stimuli.

MeSH Terms
Animals COS Cells/metabolism Calcineurin/physiology Calcium/physiology Carbachol/pharmacology Cell Line/metabolism Cell Membrane/metabolism,ultrastructure Cells, Cultured/metabolism Chlorocebus aethiops Electrophoresis, Polyacrylamide Gel Hippocampus/cytology,embryology,metabolism Humans Ion Channel Gating Ion Transport Ionomycin/pharmacology Kidney Kv1.5 Potassium Channel/chemistry,genetics,physiology Membrane Potentials Muscarinic Agonists/pharmacology Nerve Tissue Proteins/chemistry,genetics,physiology Neurons/metabolism Organ Specificity Patch-Clamp Techniques Phosphorylation Potassium/metabolism Protein Processing, Post-Translational Protein Structure, Tertiary Rats Receptors, Muscarinic/physiology Recombinant Fusion Proteins/chemistry,physiology Shab Potassium Channels/chemistry,genetics,physiology Single-Blind Method Species Specificity Structure-Activity Relationship Transfection
Chemicals
Kv1.5 Potassium Channel Muscarinic Agonists Nerve Tissue Proteins Receptors, Muscarinic Recombinant Fusion Proteins Shab Potassium Channels Ionomycin Carbachol Calcineurin Potassium Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Mohapatra Durga P
Department of Pharmacology, School of Medicine, University of California, Davis, California 95616, USA.
Trimmer James S
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Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
DOI
10.1523/JNEUROSCI.4620-05.2006
Published
2006-01-11
Pages
685-95
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6674430
Subset
IM
Grants
NCI NIH HHS · P01 CA095616 · United States
NINDS NIH HHS · R01 NS042225 · United States
NINDS NIH HHS · R01 NS042225-06 · United States
NINDS NIH HHS · NS42225 · United States
External Links
PubMed source DOI: 10.1523/JNEUROSCI.4620-05.2006 PMC Full Text View journal details
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