Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons

Alessandra Folci, Angela Steinberger, Boram Lee, Ruslan Stanika, Susanne Scheruebel, Marta Campiglio, Claudia Ramprecht, Brigitte Pelzmann, Johannes W Hell, Gerald J. Obermair, Martin Heine, Valentina Di Biase

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

L-type voltage-gated CaV1.2 calcium channels (CaV1.2) are key regulators of neuronal excitability, synaptic plasticity, and excitation-transcription coupling. Surface-exposed CaV1.2 distributes in clusters along the dendrites of hippocampal neurons. A permanent exchange between stably clustered and laterally diffusive extra-clustered channels maintains steady-state levels of CaV1.2 at dendritic signaling domains. A dynamic equilibrium between anchored and diffusive receptors is a common feature among ion channels and is crucial to modulate signaling transduction. Despite the importance of this fine regulatory system, the molecular mechanisms underlying the surface dynamics of CaV1.2 are completely unexplored. Here, we examined the dynamic states of CaV1.2 depending on phosphorylation on Ser-1700 and Ser-1928 at the channel C terminus. Phosphorylation at these sites is strongly involved in CaV1.2-mediated nuclear factor of activated T cells (NFAT) signaling, long-term potentiation, and responsiveness to adrenergic stimulation. We engineered CaV1.2 constructs mimicking phosphorylation at Ser-1700 and Ser-1928 and analyzed their behavior at the membrane by immunolabeling protocols, fluorescence recovery after photobleaching, and single particle tracking. We found that the phosphomimetic S1928E variant increases the mobility of CaV1.2 without altering the steady-state maintenance of cluster in young neurons and favors channel stabilization later in differentiation. Instead, mimicking phosphorylation at Ser-1700 promoted the diffusive state of CaV1.2 irrespective of the differentiation stage. Together, these results reveal that phosphorylation could contribute to the establishment of channel anchoring mechanisms depending on the neuronal differentiation state. Finally, our findings suggest a novel mechanism by which phosphorylation at the C terminus regulates calcium signaling by tuning the content of CaV1.2 at signaling complexes.

Original languageEnglish (US)
Pages (from-to)1040-1053
Number of pages14
JournalJournal of Biological Chemistry
Volume293
Issue number3
DOIs
StatePublished - Jan 1 2018

Fingerprint

Phosphorylation
Neurons
Fluorescence Recovery After Photobleaching
NFATC Transcription Factors
Cell signaling
Photobleaching
Neuronal Plasticity
Calcium Signaling
Long-Term Potentiation
Transcription
Dendrites
Ion Channels
Adrenergic Agents
Plasticity
L-type calcium channel alpha(1C)
Stabilization
Tuning
Fluorescence
Maintenance
Calcium

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons. / Folci, Alessandra; Steinberger, Angela; Lee, Boram; Stanika, Ruslan; Scheruebel, Susanne; Campiglio, Marta; Ramprecht, Claudia; Pelzmann, Brigitte; Hell, Johannes W; Obermair, Gerald J.; Heine, Martin; Di Biase, Valentina.

In: Journal of Biological Chemistry, Vol. 293, No. 3, 01.01.2018, p. 1040-1053.

Research output: Contribution to journalArticle

Folci, A, Steinberger, A, Lee, B, Stanika, R, Scheruebel, S, Campiglio, M, Ramprecht, C, Pelzmann, B, Hell, JW, Obermair, GJ, Heine, M & Di Biase, V 2018, 'Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons', Journal of Biological Chemistry, vol. 293, no. 3, pp. 1040-1053. https://doi.org/10.1074/jbc.M117.799585
Folci, Alessandra ; Steinberger, Angela ; Lee, Boram ; Stanika, Ruslan ; Scheruebel, Susanne ; Campiglio, Marta ; Ramprecht, Claudia ; Pelzmann, Brigitte ; Hell, Johannes W ; Obermair, Gerald J. ; Heine, Martin ; Di Biase, Valentina. / Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 3. pp. 1040-1053.
@article{fc33225b52584f70aac5bdc40a309a3d,
title = "Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons",
abstract = "L-type voltage-gated CaV1.2 calcium channels (CaV1.2) are key regulators of neuronal excitability, synaptic plasticity, and excitation-transcription coupling. Surface-exposed CaV1.2 distributes in clusters along the dendrites of hippocampal neurons. A permanent exchange between stably clustered and laterally diffusive extra-clustered channels maintains steady-state levels of CaV1.2 at dendritic signaling domains. A dynamic equilibrium between anchored and diffusive receptors is a common feature among ion channels and is crucial to modulate signaling transduction. Despite the importance of this fine regulatory system, the molecular mechanisms underlying the surface dynamics of CaV1.2 are completely unexplored. Here, we examined the dynamic states of CaV1.2 depending on phosphorylation on Ser-1700 and Ser-1928 at the channel C terminus. Phosphorylation at these sites is strongly involved in CaV1.2-mediated nuclear factor of activated T cells (NFAT) signaling, long-term potentiation, and responsiveness to adrenergic stimulation. We engineered CaV1.2 constructs mimicking phosphorylation at Ser-1700 and Ser-1928 and analyzed their behavior at the membrane by immunolabeling protocols, fluorescence recovery after photobleaching, and single particle tracking. We found that the phosphomimetic S1928E variant increases the mobility of CaV1.2 without altering the steady-state maintenance of cluster in young neurons and favors channel stabilization later in differentiation. Instead, mimicking phosphorylation at Ser-1700 promoted the diffusive state of CaV1.2 irrespective of the differentiation stage. Together, these results reveal that phosphorylation could contribute to the establishment of channel anchoring mechanisms depending on the neuronal differentiation state. Finally, our findings suggest a novel mechanism by which phosphorylation at the C terminus regulates calcium signaling by tuning the content of CaV1.2 at signaling complexes.",
author = "Alessandra Folci and Angela Steinberger and Boram Lee and Ruslan Stanika and Susanne Scheruebel and Marta Campiglio and Claudia Ramprecht and Brigitte Pelzmann and Hell, {Johannes W} and Obermair, {Gerald J.} and Martin Heine and {Di Biase}, Valentina",
year = "2018",
month = "1",
day = "1",
doi = "10.1074/jbc.M117.799585",
language = "English (US)",
volume = "293",
pages = "1040--1053",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Molecular mimicking of C-terminal phosphorylation tunes the surface dynamics of CaV1.2 calcium channels in hippocampal neurons

AU - Folci, Alessandra

AU - Steinberger, Angela

AU - Lee, Boram

AU - Stanika, Ruslan

AU - Scheruebel, Susanne

AU - Campiglio, Marta

AU - Ramprecht, Claudia

AU - Pelzmann, Brigitte

AU - Hell, Johannes W

AU - Obermair, Gerald J.

AU - Heine, Martin

AU - Di Biase, Valentina

PY - 2018/1/1

Y1 - 2018/1/1

N2 - L-type voltage-gated CaV1.2 calcium channels (CaV1.2) are key regulators of neuronal excitability, synaptic plasticity, and excitation-transcription coupling. Surface-exposed CaV1.2 distributes in clusters along the dendrites of hippocampal neurons. A permanent exchange between stably clustered and laterally diffusive extra-clustered channels maintains steady-state levels of CaV1.2 at dendritic signaling domains. A dynamic equilibrium between anchored and diffusive receptors is a common feature among ion channels and is crucial to modulate signaling transduction. Despite the importance of this fine regulatory system, the molecular mechanisms underlying the surface dynamics of CaV1.2 are completely unexplored. Here, we examined the dynamic states of CaV1.2 depending on phosphorylation on Ser-1700 and Ser-1928 at the channel C terminus. Phosphorylation at these sites is strongly involved in CaV1.2-mediated nuclear factor of activated T cells (NFAT) signaling, long-term potentiation, and responsiveness to adrenergic stimulation. We engineered CaV1.2 constructs mimicking phosphorylation at Ser-1700 and Ser-1928 and analyzed their behavior at the membrane by immunolabeling protocols, fluorescence recovery after photobleaching, and single particle tracking. We found that the phosphomimetic S1928E variant increases the mobility of CaV1.2 without altering the steady-state maintenance of cluster in young neurons and favors channel stabilization later in differentiation. Instead, mimicking phosphorylation at Ser-1700 promoted the diffusive state of CaV1.2 irrespective of the differentiation stage. Together, these results reveal that phosphorylation could contribute to the establishment of channel anchoring mechanisms depending on the neuronal differentiation state. Finally, our findings suggest a novel mechanism by which phosphorylation at the C terminus regulates calcium signaling by tuning the content of CaV1.2 at signaling complexes.

AB - L-type voltage-gated CaV1.2 calcium channels (CaV1.2) are key regulators of neuronal excitability, synaptic plasticity, and excitation-transcription coupling. Surface-exposed CaV1.2 distributes in clusters along the dendrites of hippocampal neurons. A permanent exchange between stably clustered and laterally diffusive extra-clustered channels maintains steady-state levels of CaV1.2 at dendritic signaling domains. A dynamic equilibrium between anchored and diffusive receptors is a common feature among ion channels and is crucial to modulate signaling transduction. Despite the importance of this fine regulatory system, the molecular mechanisms underlying the surface dynamics of CaV1.2 are completely unexplored. Here, we examined the dynamic states of CaV1.2 depending on phosphorylation on Ser-1700 and Ser-1928 at the channel C terminus. Phosphorylation at these sites is strongly involved in CaV1.2-mediated nuclear factor of activated T cells (NFAT) signaling, long-term potentiation, and responsiveness to adrenergic stimulation. We engineered CaV1.2 constructs mimicking phosphorylation at Ser-1700 and Ser-1928 and analyzed their behavior at the membrane by immunolabeling protocols, fluorescence recovery after photobleaching, and single particle tracking. We found that the phosphomimetic S1928E variant increases the mobility of CaV1.2 without altering the steady-state maintenance of cluster in young neurons and favors channel stabilization later in differentiation. Instead, mimicking phosphorylation at Ser-1700 promoted the diffusive state of CaV1.2 irrespective of the differentiation stage. Together, these results reveal that phosphorylation could contribute to the establishment of channel anchoring mechanisms depending on the neuronal differentiation state. Finally, our findings suggest a novel mechanism by which phosphorylation at the C terminus regulates calcium signaling by tuning the content of CaV1.2 at signaling complexes.

UR - http://www.scopus.com/inward/record.url?scp=85040996597&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85040996597&partnerID=8YFLogxK

U2 - 10.1074/jbc.M117.799585

DO - 10.1074/jbc.M117.799585

M3 - Article

C2 - 29180451

AN - SCOPUS:85040996597

VL - 293

SP - 1040

EP - 1053

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -