Phosphorylation sites on calcium channel alpha1 and beta subunits regulate ERK-dependent modulation of neuronal N-type calcium channels - PubMed

Phosphorylation sites on calcium channel alpha1 and beta subunits regulate ERK-dependent modulation of neuronal N-type calcium channels

S W Martin et al. Cell Calcium. 2006 Mar.

Abstract

Voltage-dependent calcium channels (VDCCs) in sensory neurones are tonically up-regulated via Ras/extracellular signal regulated kinase (ERK) signalling. The presence of putative ERK consensus sites within the intracellular loop linking domains I and II of neuronal N-type (Ca(v)2.2) calcium channels and all four neuronal calcium channel beta subunits (Ca(v)beta), suggests that Ca(v)2.2 and/or Ca(v)betas may be ERK-phosphorylated. Here we report that GST-Ca(v)2.2 I-II loop, and to a lesser extent Ca(v)beta1b-His(6), are substrates for ERK1/2 phosphorylation. Serine to alanine mutation of Ser-409 and/or Ser-447 on GST-Ca(v)2.2 I-II loop significantly reduced phosphorylation. Loss of Ser-447 reduced phosphorylation to a greater extent than mutation of Ser-409. Patch-clamp recordings from wild-type Ca(v)2.2,beta1b,alpha2delta1 versus mutant Ca(v)2.2(S447A) or Ca(v)2.2(S409A) channels revealed that mutation of either site significantly reduced current inhibition by UO126, a MEK (ERK kinase)-specific inhibitor that down-regulates ERK activity. However, no additive effect was observed by mutating both residues together, suggesting some functional redundancy between these sites. Mutation of both Ser-161 and Ser-348 on Ca(v)beta1b did not significantly reduce phosphorylation but did reduce UO126-induced current inhibition. Crucially, co-expression of Ca(v)2.2(S447A) with Ca(v)beta1b(S161,348A) had an additive effect, abolishing the action of UO126 on channel current, an effect not seen when Ca(v)beta1b(S161,348A) was co-expressed with Ca(v)2.2(S409A). Thus, Ser-447 on Ca(v)2.2 and Ser-161 and Ser-348 of Ca(v)beta1b appear to be both necessary and sufficient for ERK-dependent modulation of these channels. Together, our data strongly suggest that modulation of neuronal N-type VDCCs by ERK involves phosphorylation of Ca(v)2.2alpha1 and to a lesser extent possibly also Ca(v)beta subunits.

Similar articles

• Kinetics and Gbetagamma modulation of Ca(v)2.2 channels with different auxiliary beta subunits.

  Meir A, Dolphin AC. Meir A, et al. Pflugers Arch. 2002 May;444(1-2):263-75. doi: 10.1007/s00424-002-0803-3. Epub 2002 Mar 9. Pflugers Arch. 2002. PMID: 11976940

• Growth factors differentially regulate neuronal Cav channels via ERK-dependent signalling.

  Woodall AJ, Richards MA, Turner DJ, Fitzgerald EM. Woodall AJ, et al. Cell Calcium. 2008 Jun;43(6):562-75. doi: 10.1016/j.ceca.2007.10.001. Epub 2007 Nov 9. Cell Calcium. 2008. PMID: 17996937

• Functional role of a C-terminal Gbetagamma-binding domain of Ca(v)2.2 channels.

  Li B, Zhong H, Scheuer T, Catterall WA. Li B, et al. Mol Pharmacol. 2004 Sep;66(3):761-9. doi: 10.1124/mol.66.3.. Mol Pharmacol. 2004. PMID: 15322269

• Voltage-dependent calcium channels.

  Lacinová L. Lacinová L. Gen Physiol Biophys. 2005 Jun;24 Suppl 1:1-78. Gen Physiol Biophys. 2005. PMID: 16096350 Review.

• Voltage- and calcium-dependent inactivation in high voltage-gated Ca(2+) channels.

  Cens T, Rousset M, Leyris JP, Fesquet P, Charnet P. Cens T, et al. Prog Biophys Mol Biol. 2006 Jan-Apr;90(1-3):104-17. doi: 10.1016/j.pbiomolbio.2005.05.013. Epub 2005 Jul 1. Prog Biophys Mol Biol. 2006. PMID: 16038964 Review.

Cited by

• Erk1/2 inhibit synaptic vesicle exocytosis through L-type calcium channels.

  Subramanian J, Morozov A. Subramanian J, et al. J Neurosci. 2011 Mar 23;31(12):4755-64. doi: 10.1523/JNEUROSCI.6594-10.2011. J Neurosci. 2011. PMID: 21430174 Free PMC article.

• Subtype-specific reduction of voltage-gated calcium current in medium-sized dorsal root ganglion neurons after painful peripheral nerve injury.

  McCallum JB, Wu HE, Tang Q, Kwok WM, Hogan QH. McCallum JB, et al. Neuroscience. 2011 Apr 14;179:244-55. doi: 10.1016/j.neuroscience.2011.01.049. Epub 2011 Jan 28. Neuroscience. 2011. PMID: 21277351 Free PMC article.

• Interplay between electrical activity and bone morphogenetic protein signaling regulates spinal neuron differentiation.

  Swapna I, Borodinsky LN. Swapna I, et al. Proc Natl Acad Sci U S A. 2012 Oct 2;109(40):16336-41. doi: 10.1073/pnas.1202818109. Epub 2012 Sep 18. Proc Natl Acad Sci U S A. 2012. PMID: 22991474 Free PMC article.

• Apoptosis-induced alkalinization by the Na+/H+ exchanger isoform 1 is mediated through phosphorylation of amino acids Ser726 and Ser729.

  Grenier AL, Abu-ihweij K, Zhang G, Ruppert SM, Boohaker R, Slepkov ER, Pridemore K, Ren JJ, Fliegel L, Khaled AR. Grenier AL, et al. Am J Physiol Cell Physiol. 2008 Oct;295(4):C883-96. doi: 10.1152/ajpcell.00574.2007. Epub 2008 Aug 13. Am J Physiol Cell Physiol. 2008. PMID: 18701649 Free PMC article.

• Increased thrombospondin-4 after nerve injury mediates disruption of intracellular calcium signaling in primary sensory neurons.

  Guo Y, Zhang Z, Wu HE, Luo ZD, Hogan QH, Pan B. Guo Y, et al. Neuropharmacology. 2017 May 1;117:292-304. doi: 10.1016/j.neuropharm.2017.02.019. Epub 2017 Feb 22. Neuropharmacology. 2017. PMID: 28232180 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Elsevier Science

• Molecular Biology Databases

  • GlyGen glycoinformatics resource
  • PhosphoSitePlus

• Research Materials

  • NCI CPTC Antibody Characterization Program

• Miscellaneous

  • NCI CPTAC Assay Portal