Biomolecular NMR Assignments

, Volume 13, Issue 1, pp 233–237 | Cite as

Chemical shift assignments of a calmodulin intermediate with two Ca2+ bound in complex with the IQ-motif of voltage-gated Ca2+ channels (CaV1.2)

  • Ian Salveson
  • David E. Anderson
  • Johannes W. Hell
  • James B. AmesEmail author


Calcium-dependent inactivation (CDI) of neuronal voltage-gated Ca2+ channels (CaV1.2) is important for synaptic plasticity, which is associated with learning and memory. The Ca2+-dependent binding of calmodulin (CaM) to CaV1.2 is essential for CDI. Here we report NMR assignments for a CaM mutant (D21A/D23A/D25A/E32Q/D57A/D59A/N61A/E68Q, called CaMEF12) that contains two Ca2+ bound at the third and fourth EF-hands (EF3 and EF4) and is bound to the IQ-motif (residues 1644–1665) from CaV1.2 (BMRB accession no. 27692).


Calmodulin EF-hand CaV1.2 IQ-motif Synaptic plasticity CDI 



We thank Jeff Walton for technical support and help with NMR experiments. Work supported by NIH Grants (EY012347) to JBA and (AG055357) to JWH.


  1. Adams PJ, Ben-Johny M, Dick IE, Inoue T, Yue DT (2014) Apocalmodulin itself promotes ion channel opening and Ca(2+) regulation. Cell 159:608–622CrossRefGoogle Scholar
  2. Ben Johny M, Yang PS, Bazzazi H, Yue DT (2013) Dynamic switching of calmodulin interactions underlies Ca2+ regulation of CaV1.3 channels. Nat Commun 4:1717CrossRefGoogle Scholar
  3. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeiffer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293CrossRefGoogle Scholar
  4. Erickson M, Alseikhan B, Peterson B, Yue D (2001) Preassociation of calmodulin with voltage-gated Ca(2+) channels revealed by FRET in single living cells. Neuron 31:973–985CrossRefGoogle Scholar
  5. Findeisen F, Rumpf CH, Minor DL Jr (2013) Apo states of calmodulin and CaBP1 control CaV1 voltage-gated calcium channel function through direct competition for the IQ domain. J Mol Biol 425:3217–3234CrossRefGoogle Scholar
  6. Gabelli SB, Boto A, Kuhns VH, Bianchet MA, Farinelli F, Aripirala S, Yoder J, Jakoncic J, Tomaselli GF, Amzel LM (2014) Regulation of the NaV1.5 cytoplasmic domain by calmodulin. Nat Commun 5:5126ADSCrossRefGoogle Scholar
  7. Gopalakrishna R, Anderson WB (1982) Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-Sepharose affinity chromatography. Biochem Biophys Res Commun 104:830–836CrossRefGoogle Scholar
  8. Hell JW, Westenbroek RE, Warner C, Ahlijanian MK, Prystay W, Gilbert MM, Snutch TP, Catterall WA (1993) Identification and differential subcellular localization of the neuronal class C and class D L-type calcium channel alpha 1 subunits. J Cell Biol 123:949–962CrossRefGoogle Scholar
  9. Ikura M, Kay LE, Bax A (1990) A novel approach for sequential assignment of 1H, 13C, and 15N spectra of proteins: heteronuclear triple-resonance three-dimensional NMR spectroscopy. Application to calmodulin. Biochemistry 29:4659–4667CrossRefGoogle Scholar
  10. Ikura M, Spera S, Barbato G, Kay LE, Krinks M, Bax A (1991) Secondary structure and side-chain 1H and 13C resonance assignments of calmodulin in solution by heteronuclear multidimensional NMR spectroscopy. Biochemistry 30:9216–9228CrossRefGoogle Scholar
  11. Lee W, Tonelli M, Markley JL (2015) NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy. Bioinformatics 31:1325–1327CrossRefGoogle Scholar
  12. Moosmang S, Haider N, Klugbauer N, Adelsberger H, Langwieser N, Muller J, Stiess M, Marais E, Schulla V, Lacinova L, Goebbels S, Nave K, Storm D, Hofmann F, Kleppisch T (2005) Role of hippocampal Cav1.2 Ca2+ channels in NMDA receptor-independent synaptic plasticity and spatial memory. J Neurosci 25:9883–9892CrossRefGoogle Scholar
  13. Piazza M, Dieckmann T, Guillemette JG (2016) Structural studies of a complex between endothelial nitric oxide synthase and calmodulin at physiological calcium concentration. Biochemistry 55:5962–5971CrossRefGoogle Scholar
  14. Shen Y, Delaglio F, Cornilescu G, Bax A (2009) TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts. J Biomol NMR 44:213–223CrossRefGoogle Scholar
  15. Simms BA, Zamponi GW (2014) Neuronal voltage-gated calcium channels: structure, function, and dysfunction. Neuron 82:24–45CrossRefGoogle Scholar
  16. Sondergaard MT, Tian X, Liu Y, Wang R, Chazin WJ, Chen SR, Overgaard MT (2015) Arrhythmogenic calmodulin mutations affect the activation and termination of cardiac ryanodine receptor-mediated Ca2+ release. J Biol Chem 290:26151–26162CrossRefGoogle Scholar
  17. Van Petegem F, Chatelain FC, Minor DL Jr (2005) Insights into voltage-gated calcium channel regulation from the structure of the CaV1.2 IQ domain-Ca2+/calmodulin complex. Nat Struct Mol Biol 12:1108–1115CrossRefGoogle Scholar
  18. Vogl AM, Brockmann MM, Giusti SA, Maccarrone G, Vercelli CA, Bauder CA, Richter JS, Roselli F, Hafner AS, Dedic N, Wotjak CT, Vogt-Weisenhorn DM, Choquet D, Turck CW, Stein V, Deussing JM, Refojo D (2015) Neddylation inhibition impairs spine development, destabilizes synapses and deteriorates cognition. Nat Neurosci 18:239–251CrossRefGoogle Scholar
  19. Wang K, Holt C, Lu J, Brohus M, Larsen K, Overgaard M, Wimmer R, Van Petegem F (2018) Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel. Proc Natl Acad Sci USA 115:E10556–E10565CrossRefGoogle Scholar
  20. Wishart DS, Sykes BD, Richards FM (1992) The chemical shift index: a fast and simple method for the assignment of protein secondary structure through NMR spectroscopy. Biochemistry 31:1647–1651CrossRefGoogle Scholar
  21. Zuhlke RD, Pitt GS, Deisseroth K, Tsien RW, Reuter H (1999) Calmodulin supports both inactivation and facilitation of L-type calcium channels. Nature 399:159–162ADSCrossRefGoogle Scholar

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of CaliforniaDavisUSA
  2. 2.Department of PharmacologyUniversity of CaliforniaDavisUSA

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