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Interplay between R513 methylation and S516 phosphorylation of the cardiac voltage-gated sodium channel

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Abstract

Arginine methylation is a novel post-translational modification within the voltage-gated ion channel superfamily, including the cardiac sodium channel, NaV1.5. We show that NaV1.5 R513 methylation decreases S516 phosphorylation rate by 4 orders of magnitude, the first evidence of protein kinase A inhibition by arginine methylation. Reciprocally, S516 phosphorylation blocks R513 methylation. NaV1.5 p.G514C, associated to cardiac conduction disease, abrogates R513 methylation, while leaving S516 phosphorylation rate unchanged. This is the first report of methylation–phosphorylation cross-talk of a cardiac ion channel.

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Fig. 1

Abbreviations

MALDI-TOF:

Matrix-assisted laser desorption–ionization time of flight

NaV1.5:

Voltage-gated sodium channel, cardiac isoform, α subunit

PKA:

cAMP-dependent protein kinase

PRMT:

Protein arginine methyltransferase

SAM:

S-adenosyl-l-methionine

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Acknowledgments

The clone for expression of PRMT3 as GST fusion was a gift from Alexsandra Espejo and Mark. T. Bedford (MD Anderson Cancer Center). We thank Ariel Escobar (UC Merced), Guillermo Pérez, and Fabiana Scornik (University of Girona) for critical reading of the manuscript. Peptide synthesis was performed by Javier Valle and David Andreu (Laboratory of Proteomics and Protein Chemistry, University Pompeu Fabra, Barcelona). We thank Antonia Odena and Eliandre de Oliveira (Proteomics Platform, University of Barcelona, member of the ProteoRed-ISCIII network), and Guadalupe Espadas, Jenny G. Donoso and Eduard Sabidó (CRG/UPF Proteomics Unit) for help with LC–MS/MS analyses. This work was funded by Fundació Obra Social La Caixa, Spanish Government [SAF2011-27627, CTQ2011-25086/BQU)], and European Community [PCIG14-GA-2013-630978]. PB-A acknowledges a Sara Borrell fellowship [CD10/00275], SO a JdC contract [JCI-2012-14438] and FF a Beatriu de Pinós fellowship [BP-2010-A2_00022].

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Cardiovascular Genetics Center, Institut d’Investigació Biomèdica de Girona Dr. Josep Trueta, University of Girona, 17003, Girona, Spain

    Pedro Beltran-Alvarez, Rubí Díaz-Hernández, Ramon Brugada & Sara Pagans

  2. Department of Medical Sciences, School of Medicine, University of Girona, 17003, Girona, Spain

    Pedro Beltran-Alvarez, Rubí Díaz-Hernández, Ramon Brugada & Sara Pagans

  3. Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona, Campus Montilivi, 17071, Girona, Spain

    Ferran Feixas & Sílvia Osuna

Authors
  1. Pedro Beltran-Alvarez
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  2. Ferran Feixas
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  3. Sílvia Osuna
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  4. Rubí Díaz-Hernández
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  5. Ramon Brugada
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  6. Sara Pagans
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Corresponding authors

Correspondence to Pedro Beltran-Alvarez, Ramon Brugada or Sara Pagans.

Additional information

F. Feixas and S. Osuna contributed equally.

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Beltran-Alvarez, P., Feixas, F., Osuna, S. et al. Interplay between R513 methylation and S516 phosphorylation of the cardiac voltage-gated sodium channel. Amino Acids 47, 429–434 (2015). https://doi.org/10.1007/s00726-014-1890-0

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  • DOI: https://doi.org/10.1007/s00726-014-1890-0

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