Critical Role of cAMP-Dependent Protein Kinase Anchoring to the L-Type Calcium Channel Ca<sub>v</sub>1.2 via A-Kinase Anchor Protein 150 in Neurons

Hall, Duane D.; Davare, Monika A.; Shi, Mei; Allen, Margaret L.; Weisenhaus, Michael; McKnight, G. Stanley; Hell, Johannes

doi:10.1021/bi062217x

Cited by 123 publications

(168 citation statements)

References 84 publications

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“…Although we observed Yotiao, another abundant neuro- nal AKAP, in our lysates, we did not find that it co-immunoprecipitated with Ca V 1.2 channels, further verifying the specificity of this interaction. These results confirm the identification of AKAP15 in these co-immunoprecipitates as well as ␤ 2 ARs, MAP2B, and AKAP150 as reported previously (28,39). We have previously found that AKAP15 binds directly to cardiac Ca V 1.2 channels via the C-terminal LZ motif (24), so it is likely that the co-immunoprecipitation observed here reflects direct interaction between these two proteins.…”

Section: Resultssupporting

confidence: 92%

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

Marshall

Clark

Westenbroek

et al. 2011

Journal of Biological Chemistry

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Regulation of Ca V 1.2 channels in cardiac myocytes by the ␤-adrenergic pathway requires a signaling complex in which the proteolytically processed distal C-terminal domain acts as an autoinhibitor of channel activity and mediates up-regulation by the ␤-adrenergic receptor and PKA bound to A-kinase anchoring protein 15 (AKAP15). We examined the significance of this distal C-terminal signaling complex for Ca V 1.2 and Ca V 1.3 channels in neurons. AKAP15 co-immunoprecipitates with Ca V 1.2 and Ca V 1.3 channels. AKAP15 has overlapping localization with Ca V 1.2 and Ca V 1.3 channels in cell bodies and proximal dendrites and is closely co-localized with Ca V 1.2 channels in punctate clusters. The neuronal AKAP MAP2B, which also interacts with Ca V 1.2 and Ca V 1.3 channels, has complementary localization to AKAP15, suggesting different functional roles in calcium channel regulation. Studies with mice that lack the distal C-terminal domain of Ca V 1.2 channels (Ca V 1.2⌬DCT) reveal that AKAP15 interacts with neuronal Ca V 1.2 channels via their C terminus in vivo and is co-localized in punctate clusters of Ca V 1.2 channels via that interaction. Ca V 1.2⌬DCT neurons have reduced L-type calcium current, indicating that the distal C-terminal domain is required for normal functional expression in vivo. Deletion of the distal C-terminal domain impairs calcium-dependent signaling from Ca V 1.2 channels to the nucleus, as shown by reduction in phosphorylation of the cAMP response element-binding protein. Our results define AKAP signaling complexes of Ca V 1.2 and Ca V 1.3 channels in brain and reveal three previously unrecognized functional roles for the distal C terminus of neuronal Ca V 1.2 channels in vivo: increased functional expression, anchoring of AKAP15 and PKA, and initiation of excitation-transcription coupling.Voltage-gated calcium channels of the Ca V 1 subfamily conduct L-type calcium currents that transduce cell-surface depolarization into calcium transients and initiate excitation-contraction coupling, excitation-secretion coupling, protein phosphorylation, and gene regulation (1-5). Calcium influx via postsynaptic Ca V 1 channels supports sustained phosphorylation of cAMP response element-binding protein (CREB) 3 and CREB-dependent gene expression in hippocampal neurons (6 -13).Functional Ca V 1 channels are multimeric complexes composed of pore-forming ␣ 1 and associated ␣ 2 ␦, ␤, and in some cases, ␥ subunits (14 -19). These channels have an extended C terminus containing many protein interaction sites for regulation (5). In brain, Ca V 1 channels are composed of 70% Ca V 1.2 and 22% Ca V 1.3 with minor contributions from other Ca V 1 channels, as indicated by immunoprecipitation with specific antibodies (20). Ca V 1.2 and Ca V 1.3 channels are primarily localized in the soma and proximal dendrites (20, 21).The ␤-adrenergic pathway activates cAMP-dependent protein kinase (PKA) and increases the activity of Ca V 1 channels in skeletal and cardiac myocytes and neurons (1-5, 22, 23). PKAmediated regulatio...

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Section: Resultssupporting

confidence: 92%

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

Marshall

Clark

Westenbroek

et al. 2011

Journal of Biological Chemistry

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“…AKAP150 KO and D36 mouse strains were generated as previously described (22,25,52). Briefly, the entire coding sequence of the AKAP150 (Akap5) gene (GenBank TM locus XM138063 position 2126 -2131) was replaced with a neomycin phosphotransferase cassette in AKAP150 KO mice by homologous recombination in embryonic stem cells in a 129S1/SvImJ background (Jackson Laboratories, Bar Harbor, ME).…”

Section: Methodsmentioning

confidence: 99%

“…We previously generated AKAP150 KO mice and mice that lack the PKA binding site at the C terminus of AKAP150 (D36 mice) (22,25,52). We showed earlier that PKA and its anchoring by AKAP150 plays an important role in LTP at an age of 2-3 weeks and again from 8 weeks on but not during 3-6 weeks (22).…”

mentioning

confidence: 99%

A Kinase Anchor Protein 150 (AKAP150)-associated Protein Kinase A Limits Dendritic Spine Density

Lü

Zha

Kim

et al. 2011

Journal of Biological Chemistry

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The A kinase anchor protein AKAP150 recruits the cAMPdependent protein kinase (PKA) to dendritic spines. Here we show that in AKAP150 (AKAP5) knock-out (KO) mice frequency of miniature excitatory post-synaptic currents (mEPSC) and inhibitory post-synaptic currents (mIPSC) are elevated at 2 weeks and, more modestly, 4 weeks of age in the hippocampal CA1 area versus litter mate WT mice. Linear spine density and ratio of AMPAR to NMDAR EPSC amplitudes were also increased. Amplitude and decay time of mEPSCs, decay time of mIPSCs, and spine size were unaltered. Mice in which the PKA anchoring C-terminal 36 residues of AKAP150 are deleted (D36) showed similar changes. Furthermore, whereas acute stimulation of PKA (2-4 h) increases spine density, prolonged PKA stimulation (48 h) reduces spine density in apical dendrites of CA1 pyramidal neurons in organotypic slice cultures. The data from the AKAP150 mutant mice show that AKAP150-anchored PKA chronically limits the number of spines with functional AMPARs at 2-4 weeks of age. However, synaptic transmission and spine density was normal at 8 weeks in KO and D36 mice. Thus AKAP150-independent mechanisms correct the aberrantly high number of active spines in juvenile AKAP150 KO and D36 mice during development.Dendritic spines receive most of the excitatory input in mammalian neurons (1, 2) and compartmentalize signaling that regulates the synaptic response (3, 4). Several protein kinases and phosphatases, including CaMKII, cAMP-dependent protein kinase (PKA), 3 PKC, and protein phosphatase PP2A and PP2B (calcineurin) (5) play prominent roles in synaptic plasticity. Synaptic activity translates into their activation, thereby affecting spine stability, morphology, and postsynaptic responses (6 -13). Spines are dynamically gained and lost during synaptogenesis (9, 14 -16) and in response to neuronal activity (17-19).AKAP150 is a product of the Akap5 gene in mice and is the main AKAP that recruits PKA to postsynaptic sites (20 -25). It binds with its very C terminus to the N-terminal dimerization domain of the regulatory RII subunits of PKA (26,27). It also binds PKC (28), PP2B (29,30), and the SH3-GK region of the postsynaptic scaffolding proteins SAP97 and PSD-95 (31, 32). PSD-95 binds with its PDZ domains to the C-terminal ((S/ T)X(I/V)) motif of NMDA-type glutamate receptor (NMDAR) NR2 subunits (33). PSD-95 also binds to the C terminus of stargazin (␥2) and its homologues ␥3, ␥4, and ␥8, which associate with AMPAR-type glutamate receptor (AMPAR) for postsynaptic localization in conjunction with . SAP97 directly binds to the AMPAR GluR1 subunit via a PDZ-C-terminal interaction (37) to link AKAP150 and thereby PKA, PKC,. Phosphorylation of GluR1 on Ser-845 by PKA can increase channel activity (42) and accumulation of GluR1-containing AMPAR at the postsynaptic site during hippocampal LTP (43, 44) (see also Refs. 45,46). The N terminus of AKAP150 also binds F-actin, cadherin, adenylyl cyclases, and PIP 2 and targets AKAP150 to dendritic spines and the central region binds PSD-...

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“…Previous studies showed that AKAP79/150 is abundant at the postsynaptic sites of glutamatergic synapses in which it facilitates PKA-dependent phosphorylation of AMPA and NMDA glutamate receptors, thereby contributing to synaptic plasticity (Colledge et al, 2000;Gomez et al, 2002;Lu et al, 2007). AKAP79/150 also associates with the L-type (Ca v 1.2) Ca 2ϩ channels (Gao et al, 1997;Hall et al, 2007), M-type (KCNQ2/3) K ϩ channels (Hoshi et al, 2003), ASIC1a and 2a channels (Chai et al, 2007), and ␤2-adrenergic receptor (Fraser et al, 2000).…”

Section: Trpv1 and Akap150 Are Coexpressed And Coimmunoprecipitate Inmentioning

confidence: 99%

“…We studied expression of four different AKAPs that have been shown to associate with other ion channels and receptors: AKAP15/18, AKAP150, AKAP250, and Yotiao (Wong and Scott, 2004). Immunoblots obtained from DRG and spinal cord lysates were compared with those obtained from brain, in which these AKAPs have been characterized previously (Tibbs et al, 1998;Westphal et al, 1999;Colledge et al, 2000;Piontek and Brandt, 2003;Hall et al, 2007;Lu et al, 2007).…”

Section: Trpv1 and Akap150 Are Coexpressed And Coimmunoprecipitate Inmentioning

confidence: 99%

Protein Kinase A Anchoring via AKAP150 Is Essential for TRPV1 Modulation by Forskolin and Prostaglandin E₂in Mouse Sensory Neurons

Schnizler¹,

Shutov²,

Kanegan³

et al. 2008

J. Neurosci.

161

174

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Phosphorylation-dependent modulation of the vanilloid receptor TRPV1 is one of the key mechanisms mediating the hyperalgesic effects of inflammatory mediators, such as prostaglandin E 2 (PGE 2 ). However, little is known about the molecular organization of the TRPV1 phosphorylationcomplexandspecificallyaboutscaffoldingproteinsthatpositiontheproteinkinaseA(PKA)holoenzymeproximaltoTRPV1foreffective and selective regulation of the receptor. Here, we demonstrate the critical role of the A-kinase anchoring protein AKAP150 in PKA-dependent modulation of TRPV1 function in adult mouse dorsal root ganglion (DRG) neurons. We found that AKAP150 is expressed in ϳ80% of TRPV1-positive DRG neurons and is coimmunoprecipitated with the capsaicin receptor. In functional studies, PKA stimulation with forskolin markedly reduced desensitization of TRPV1. This effect was blocked by the PKA selective inhibitors KT5720 [(9S,10R,12R)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-1-oxo-9, 12-epoxy-1H-diindolo[1,2,3-fg:3Ј,2Ј,1Ј-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylicacid hexyl ester] and H89 (N-[2-( p-bromo-cinnamylamino)-ethyl]-5-isoquinoline-sulfon-amide 2HCl), as well as by the AKAP inhibitory peptide Ht31. Similarly, PGE 2 decreased TRPV1 desensitization in a manner sensitive to the PKA inhibitor KT5720. Both the forskolin and PGE 2 effects were strongly impaired in DRG neurons from knock-in mice that express a mutant AKAP150 lacking the PKA-binding domain (⌬36 mice). Protein kinase C-dependent sensitization of TRPV1 remained intact in ⌬36 mice. The PGE 2 /PKA signaling defect in DRG neurons from ⌬36 mice was rescued by overexpressing the full-length human ortholog of AKAP150 in these cells. In behavioral testing, PGE 2 -induced thermal hyperalgesia was significantly diminished in ⌬36 mice. Together, these data suggest that PKA anchoring by AKAP150 is essential for the enhancement of TRPV1 function by activation of the PGE 2 /PKA signaling pathway.

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Critical Role of cAMP-Dependent Protein Kinase Anchoring to the L-Type Calcium Channel Ca_v1.2 via A-Kinase Anchor Protein 150 in Neurons

Cited by 123 publications

References 84 publications

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

A Kinase Anchor Protein 150 (AKAP150)-associated Protein Kinase A Limits Dendritic Spine Density

Protein Kinase A Anchoring via AKAP150 Is Essential for TRPV1 Modulation by Forskolin and Prostaglandin E₂in Mouse Sensory Neurons

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Critical Role of cAMP-Dependent Protein Kinase Anchoring to the L-Type Calcium Channel Cav1.2 via A-Kinase Anchor Protein 150 in Neurons

Cited by 123 publications

References 84 publications

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

Functional Roles of a C-terminal Signaling Complex of CaV1 Channels and A-kinase Anchoring Protein 15 in Brain Neurons

A Kinase Anchor Protein 150 (AKAP150)-associated Protein Kinase A Limits Dendritic Spine Density

Protein Kinase A Anchoring via AKAP150 Is Essential for TRPV1 Modulation by Forskolin and Prostaglandin E2in Mouse Sensory Neurons

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Critical Role of cAMP-Dependent Protein Kinase Anchoring to the L-Type Calcium Channel Ca_v1.2 via A-Kinase Anchor Protein 150 in Neurons

Protein Kinase A Anchoring via AKAP150 Is Essential for TRPV1 Modulation by Forskolin and Prostaglandin E₂in Mouse Sensory Neurons