M-type (KCNQ2/3) potassium channels are suppressed by activation of Gq/11-coupled receptors, thereby increasing neuronal excitability. We show here that rat KCNQ2 can bind directly to the multivalent A-kinase-anchoring protein AKAP150. Peptides that block AKAP150 binding to the KCNQ2 channel complex antagonize the muscarinic inhibition of the currents. A mutant form of AKAP150, AKAP(ΔA), which is unable to bind protein kinase C (PKC), also attenuates the agonist-induced current suppression. Analysis of recombinant KCNQ2 channels suggests that targeting of PKC through association with AKAP150 is important for the inhibition. Phosphorylation of KCNQ2 channels was increased by muscarinic stimulation; this was prevented either by coexpression with AKAP(ΔA) or pretreatment with PKC inhibitors that compete with diacylglycerol. These inhibitors also reduced muscarinic inhibition of M-current. Our data indicate that AKAP150-bound PKC participates in receptor-induced inhibition of the M-current.
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Brown, D.A. M-current. in Ion Channels (ed. Narahashi, T.) 55–94 (Plenum, New York, 1988).
Marrion, N.V. Control of M-current. Annu. Rev. Physiol. 59, 483–504 (1997).
Aiken, S.P., Lampe, B.J., Murphy, P.A. & Brown, B.S. Reduction of spike frequency adaptation and blockade of M-current in rat CA1 pyramidal neurones by linopirdine (DuP 996), a neurotransmitter release enhancer. Br. J. Pharmacol. 115, 1163–1168 (1995).
Lamas, J.A., Selyanko, A.A. & Brown, D.A. Effects of a cognition-enhancer, linopirdine (DuP 996), on M-type potassium currents (IK(M)) and some other voltage- and ligand-gated membrane currents in rat sympathetic neurons. Eur. J. Neurosci. 9, 605–616 (1997).
Rundfeldt, C. & Netzer, R. The novel anticonvulsant retigabine activates M-currents in Chinese hamster ovary cells tranfected with human KCNQ2/3 subunits. Neurosci. Lett. 282, 73–76 (2000).
Tatulian, L., Delmas, P., Abogadie, F.C. & Brown, D.A. Activation of expressed KCNQ potassium currents and native neuronal M-type potassium currents by the anti-convulsant drug retigabine. J. Neurosci. 21, 5535–5545 (2001).
Wang, H.S. et al. KCNQ2 and KCNQ3 potassium channel subunits: molecular correlates of the M-channel. Science 282, 1890–1893 (1998).
Selyanko, A.A. et al. Inhibition of KCNQ1-4 potassium channels expressed in mammalian cells via M1 muscarinic acetylcholine receptors. J. Physiol. 522, 349–355 (2000).
Shapiro, M.S. et al. Reconstitution of muscarinic modulation of the KCNQ2/KCNQ3 K+ channels that underlie the neuronal M-current. J. Neurosci. 20, 1710–1721 (2000).
Jentsch, T.J. Neuronal KCNQ potassium channels: physiology and role in disease. Nat. Rev. Neurosci. 1, 21–30 (2000).
Brown, B.S. & Yu, S.P. Modulation and genetic identification of the M-channel. Prog. Biophys. Mol. Biol. 73, 135–166 (2000).
Haley, J.E. et al. The alpha subunit of Gq contributes to muscarinic inhibition of the M-type potassium current in sympathetic neurons. J. Neurosci. 18, 4521–4531 (1998).
Haley, J.E. et al. Bradykinin, but not muscarinic, inhibition of M-current in rat sympathetic ganglion neurons involves phospholipase C-beta 4. J. Neurosci. 20, RC105 (2000).
Selyanko, A.A., Stansfeld, C.E. & Brown, D.A. Closure of potassium M-channels by muscarinic acetylcholine-receptor stimulants requires a diffusible messenger. Proc. R. Soc. Lond. B Biol. Sci. 250, 119–125 (1992).
Marrion, N.V. M-current suppression by agonist and phorbol ester in bullfrog sympathetic neurons. Pflugers. Arch. 426, 296–303 (1994).
Selyanko, A.A. & Brown, D.A. Intracellular calcium directly inhibits potassium M channels in excised membrane patches from rat sympathetic neurons. Neuron 16, 151–162 (1996).
Cruzblanca, H., Koh, D.S. & Hille, B. Bradykinin inhibits M current via phospholipase C and Ca2+ release from IP3-sensitive Ca2+ stores in rat sympathetic neurons. Proc. Natl. Acad. Sci. USA 95, 7151–7156 (1998).
Bofill-Cardona, E. et al. Two different signaling mechanisms involved in the excitation of rat sympathetic neurons by uridine nucleotides. Mol. Pharmacol. 57, 1165–1172 (2000).
Robbins, J., Marsh, S.J. & Brown, D.A. On the mechanism of M-current inhibition by muscarinic m1 receptors in DNA-transfected rodent neuroblastoma × glioma cells. J. Physiol. 469, 153–178 (1993).
del Rio, E. et al. Muscarinic M1 receptors activate phosphoinositide turnover and Ca2+ mobilisation in rat sympathetic neurones, but this signalling pathway does not mediate M-current inhibition. J. Physiol. 520, 101–111 (1999).
Suh, B.C. & Hille, B. Recovery from muscarinic modulation of M-current channels requires phosphatidylinositol 4,5-bisphosphate synthesis. Neuron 35, 507–520 (2002).
Marrion, N.V. Calcineurin regulates M channel modal gating in sympathetic neurons. Neuron 16, 163–173 (1996).
Higashida, H. & Brown, D.A. Two polyphosphatidylinositide metabolites control two K+ currents in a neuronal cell. Nature 323, 333–335 (1986).
Pawson, T. & Scott, J.D. Signaling through scaffold, anchoring, and adaptor proteins. Science 278, 2075–2080 (1997).
Sudol, M. & Hunter, T. NeW wrinkles for an old domain. Cell 103, 1001–1004 (2000).
Colledge, M. & Scott, J.D. AKAPs: from structure to function. Trends Cell Biol. 9, 216–221 (1999).
Fraser, I.D., Scott, J.D. Modulation of ion channels: a “current” view of AKAPs. Neuron 23, 423–426 (1999).
Westphal, R.S. et al. Regulation of NMDA receptors by an associated phosphatase-kinase signaling complex. Science 285, 93–96 (1999).
Coghlan, V.M. et al. Association of protein kinase A and protein phosphatase 2B with a common anchoring protein. Science 267, 108–111 (1995).
Klauck, T.M. et al. Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science 271, 1589–1592 (1996).
Colledge, M. et al. Targeting of PKA to glutamate receptors through a MAGUK-AKAP complex. Neuron 27, 107–119 (2000).
Wen, H. & Levitan, I.B. Calmodulin is an auxiliary subunit of KCNQ2/3 potassium channels. J. Neurosci. 22, 7991–8001 (2002).
Dorje, F. et al. Antagonist binding profiles of five cloned human muscarinic receptor subtypes. J. Pharmacol. Exp. Ther. 256, 727–733 (1991).
Dell'Acqua, M.L. et al. Membrane-targeting sequences on AKAP79 bind phosphatidylinositol-4, 5-bisphosphate. EMBO J. 17, 2246–2260 (1998).
Faux, M.C. et al. Mechanism of A-kinase-anchoring protein 79 (AKAP79) and protein kinase C interaction. Biochem. J. 343, 443–452 (1999).
Toullec, D. et al. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J. Biol. Chem. 266, 15771–15781 (1991).
Herbert, J.M., Augereau, J.M., Gleye, J. & Maffrand, J.P. Chelerythrine is a potent and specific inhibitor of protein kinase C. Biochem. Biophys. Res. Commun. 172, 993–999 (1990).
Kobayashi, E., Nakano, H., Morimoto, M. & Tamaoki, T. Calphostin C (UCN-1028C), a novel microbial compound, is a highly potent and specific inhibitor of protein kinase C. Biochem. Biophys. Res. Commun. 159, 548–553 (1989).
Sachs, C.W., Safa, A.R., Harrison, S.D. & Fine, R.L. Partial inhibition of multidrug resistance by safingol is independent of modulation of P-glycoprotein substrate activities and correlated with inhibition of protein kinase C. J. Biol. Chem. 270, 26639–26648 (1995).
Rosenmund, C. et al. Anchoring of protein kinase A is required for modulation of AMPA/kainate receptors on hippocampal neurons. Nature 368, 853–856 (1994).
Tavalin, S.J. et al. Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression. J. Neurosci. 22, 3044–3051 (2002).
Johnson, B.D., Scheuer, T. & Catterall, W.A. Voltage-dependent potentiation of L-type Ca2+ channels in skeletal muscle cells requires anchored cAMP-dependent protein kinase. Proc. Natl. Acad. Sci. USA 91, 11492–11496 (1994).
Gao, T. et al. cAMP-dependent regulation of cardiac L-type Ca2+ channels requires membrane targeting of PKA and phosphorylation of channel subunits. Neuron 19, 185–196 (1997).
Cooper, E.C. et al. Colocalization and coassembly of two human brain M-type potassium channel subunits that are mutated in epilepsy. Proc. Natl. Acad. Sci. USA 97, 4914–4919 (2000).
Sik, A. et al. Localization of the A kinase anchoring protein AKAP79 in the human hippocampus. Eur. J. Neurosci. 12, 1155–1164 (2000).
Marx, S.O. et al. Requirement of a macromolecular signaling complex for beta adrenergic receptor modulation of the KCNQ1-KCNE1 potassium channel. Science 295, 496–499 (2002).
Delmas, P. et al. Signaling microdomains define the specificity of receptor-mediated InsP(3) pathways in neurons. Neuron 34, 209–220 (2002).
Delmas, P. et al. On the role of endogenous G-protein beta gamma subunits in N-type Ca2+ current inhibition by neurotransmitters in rat sympathetic neurones. J. Physiol. 506, 319–329 (1998).
Takahashi, Y. et al. 12-Lipoxygenase overexpression in rodent NG108-15 cells enhances membrane excitability by inhibiting M-type K+ channels. J. Physiol. 521, 567–574 (1999).
Hoshi, N. et al. KCR1, a membrane protein that facilitates functional expression of non-inactivating K+ currents associates with rat EAG voltage-dependent K+ channels. J. Biol. Chem. 273, 23080–23085 (1998).
The authors thank M. Okamura, T. Haga and T. I. Bonner for the gift of CHO hm1 cells, and T. Rafiq for help with ganglion cell cultures. This work was supported by grants to N.H. and H.H. from the Japanese Ministry of Education, Culture, Sports, Science and Technology, by grant PG7909913 to D.A.B. from the UK Medical Research Council and by National Institute of Health grant GM48231 for the support of J.D.S.
The authors declare no competing financial interests.
Mapping the KCNQ2 binding domain within residue 1-143 of AKAP150. (a) AKAP150 interacts with KCNQ2 through multi-site contacts. Schematic diagram showing the AKAP150(1-143) deletion constructs. (b) Interaction with AKAP150 fragments were detected using the KCNQ2(321-499) overlay assay as in Fig. 2e (upper panel). Commassie stain indicates approximately equal protein loading (lower panel). (c) The amalgamation of four independent experiments is shown. (JPG 33 kb)
About this article
Cite this article
Hoshi, N., Zhang, JS., Omaki, M. et al. AKAP150 signaling complex promotes suppression of the M-current by muscarinic agonists. Nat Neurosci 6, 564–571 (2003). https://doi.org/10.1038/nn1062
RSC Medicinal Chemistry (2021)
Trends in Pharmacological Sciences (2020)
Pflügers Archiv - European Journal of Physiology (2020)
Histone deacetylase inhibition reduces ventral tegmental area dopamine neuronal hyperexcitability involving AKAP150 signaling following maternal deprivation in juvenile male rats
Journal of Neuroscience Research (2020)
Frontiers in Physiology (2020)