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The Cav1.2 N terminus contains a CaM kinase site that modulates channel trafficking and function

  • Ion channels, receptors and transporters
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Abstract

The L-type voltage-gated calcium channel Cav1.2 and the calcium-activated CaM kinase cascade both regulate excitation transcription coupling in the brain. CaM kinase is known to associate with the C terminus of Cav1.2 in a region called the PreIQ-IQ domain, which also binds multiple calmodulin molecules. Here we identify and characterize a second CaMKII binding site in the N terminus of Cav1.2 that is formed by a stretch of four amino residues (cysteine–isoleucine–serine–isoleucine) and which regulates channel expression and function. By using live cell imaging of tsA-201 cells we show that GFP fusion constructs of the CaMKII binding region, termed N2B-II co-localize with mCherry-CaMKII. Mutating CISI to AAAA ablates binding to and colocalization with CaMKII. Cav1.2-AAAA channels show reduced cell surface expression in tsA-201 cells, but interestingly, display an increase in channel function that offsets the trafficking deficit. Altogether our data reveal that the proximal N terminus of Cav1.2 contains a CaMKII binding region which contributes to channel surface expression and function.

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Abbreviations

CaMK:

Calmodulin kinase

CaMKK:

Calmodulin kinase kinase

CaM:

Calmodulin

CDF:

Calcium-dependent facilitation

CDI:

Calcium-dependent inactivation

VDF:

Voltage-dependent facilitation

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Acknowledgments

This work was supported by a grant from the Natural Sciences and Engineering Research Council. BAS is supported by a studentship from Alberta Innovates-Health Solutions (AI-HS). IAS is supported by a postdoctoral fellowship from Mitacs Elevate. GWZ is an AI-HS Scientist and a Canada Research Chair.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Standards

All experiments performed in this manuscript comply with the laws of Canada.

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Hotchkiss Brain Institute, University of Calgary, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada

    Brett A. Simms, Ivana A. Souza, Renata Rehak & Gerald W. Zamponi

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  1. Brett A. Simms
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  2. Ivana A. Souza
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  3. Renata Rehak
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  4. Gerald W. Zamponi
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Corresponding author

Correspondence to Gerald W. Zamponi.

Additional information

B.A. Simms and I.A. Souza contributed equally to this study.

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Simms, B.A., Souza, I.A., Rehak, R. et al. The Cav1.2 N terminus contains a CaM kinase site that modulates channel trafficking and function. Pflugers Arch - Eur J Physiol 467, 677–686 (2015). https://doi.org/10.1007/s00424-014-1538-7

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