The CaMKII Holoenzyme Structure in Activation-competent Conformations

Overview

Journal Nat Commun

Specialty Biology

Date 2017 Jun 8

PMID 28589927

Citations 64

Authors

Janette B Myers

Vincent Zaegel

Steven J Coultrap

Adam P Miller

K Ulrich Bayer

Steve L Reichow

Affiliations

Soon will be listed here.

Abstract

The Ca/calmodulin-dependent protein kinase II (CaMKII) assembles into large 12-meric holoenzymes, which is thought to enable regulatory processes required for synaptic plasticity underlying learning, memory and cognition. Here we used single particle electron microscopy (EM) to determine a pseudoatomic model of the CaMKIIα holoenzyme in an extended and activation-competent conformation. The holoenzyme is organized by a rigid central hub complex, while positioning of the kinase domains is highly flexible, revealing dynamic holoenzymes ranging from 15-35 nm in diameter. While most kinase domains are ordered independently, ∼20% appear to form dimers and <3% are consistent with a compact conformation. An additional level of plasticity is revealed by a small fraction of bona-fide 14-mers (<4%) that may enable subunit exchange. Biochemical and cellular FRET studies confirm that the extended state of CaMKIIα resolved by EM is the predominant form of the holoenzyme, even under molecular crowding conditions.

Citing Articles

Calcium induced N-terminal gating and pore collapse in connexin-46/50 gap junctions.

Flores J, ONeill S, Jarodsky J, Reichow S bioRxiv. 2025; .

PMID: 39990482 PMC: 11844560. DOI: 10.1101/2025.02.12.637955.

Oxidation of CaMKIIα cysteines inhibits autonomous activation induced by phosphorylation.

Rocco-Machado N, Deng M, He Y, Levine R Arch Biochem Biophys. 2024; 764():110268.

PMID: 39674564 PMC: 11750588. DOI: 10.1016/j.abb.2024.110268.

A revised view of the role of CaMKII in learning and memory.

Bayer K, Giese K Nat Neurosci. 2024; 28(1):24-34.

PMID: 39558039 DOI: 10.1038/s41593-024-01809-x.

Ligand-induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity.

Narayanan D, Larsen A, Gauger S, Adafia R, Hammershoi R, Hamborg L Protein Sci. 2024; 33(10):e5152.

PMID: 39275999 PMC: 11400628. DOI: 10.1002/pro.5152.

Hub stability in the calcium calmodulin-dependent protein kinase II.

Chien C, Puhl H, Vogel S, Molloy J, Chiu W, Khan S Commun Biol. 2024; 7(1):766.

PMID: 38918547 PMC: 11199487. DOI: 10.1038/s42003-024-06423-y.

References

Meador W, Means A, Quiocho F . Modulation of calmodulin plasticity in molecular recognition on the basis of x-ray structures. Science. 1993; 262(5140):1718-21. DOI: 10.1126/science.8259515. View

OLeary H, Lasda E, Bayer K . CaMKIIbeta association with the actin cytoskeleton is regulated by alternative splicing. Mol Biol Cell. 2006; 17(11):4656-65. PMC: 1635389. DOI: 10.1091/mbc.e06-03-0252. View

Bayer K, Lebel E, McDonald G, OLeary H, Schulman H, De Koninck P . Transition from reversible to persistent binding of CaMKII to postsynaptic sites and NR2B. J Neurosci. 2006; 26(4):1164-74. PMC: 2890238. DOI: 10.1523/JNEUROSCI.3116-05.2006. View

Goodell D, Eliseeva T, Coultrap S, Bayer K . CaMKII binding to GluN2B is differentially affected by macromolecular crowding reagents. PLoS One. 2014; 9(5):e96522. PMC: 4010494. DOI: 10.1371/journal.pone.0096522. View

Hoffman L, Stein R, Colbran R, Mchaourab H . Conformational changes underlying calcium/calmodulin-dependent protein kinase II activation. EMBO J. 2011; 30(7):1251-62. PMC: 3094114. DOI: 10.1038/emboj.2011.40. View

Coultrap S, Freund R, OLeary H, Sanderson J, Roche K, DellAcqua M . Autonomous CaMKII mediates both LTP and LTD using a mechanism for differential substrate site selection. Cell Rep. 2014; 6(3):431-7. PMC: 3930569. DOI: 10.1016/j.celrep.2014.01.005. View

Coultrap S, Barcomb K, Bayer K . A significant but rather mild contribution of T286 autophosphorylation to Ca2+/CaM-stimulated CaMKII activity. PLoS One. 2012; 7(5):e37176. PMC: 3353915. DOI: 10.1371/journal.pone.0037176. View

Hanson P, Meyer T, Stryer L, Schulman H . Dual role of calmodulin in autophosphorylation of multifunctional CaM kinase may underlie decoding of calcium signals. Neuron. 1994; 12(5):943-56. DOI: 10.1016/0896-6273(94)90306-9. View

Scheres S . RELION: implementation of a Bayesian approach to cryo-EM structure determination. J Struct Biol. 2012; 180(3):519-30. PMC: 3690530. DOI: 10.1016/j.jsb.2012.09.006. View

10.

Drozdetskiy A, Cole C, Procter J, Barton G . JPred4: a protein secondary structure prediction server. Nucleic Acids Res. 2015; 43(W1):W389-94. PMC: 4489285. DOI: 10.1093/nar/gkv332. View

11.

De Koninck P, Schulman H . Sensitivity of CaM kinase II to the frequency of Ca2+ oscillations. Science. 1998; 279(5348):227-30. DOI: 10.1126/science.279.5348.227. View

12.

Singla S, Hudmon A, Goldberg J, Smith J, Schulman H . Molecular characterization of calmodulin trapping by calcium/calmodulin-dependent protein kinase II. J Biol Chem. 2001; 276(31):29353-60. DOI: 10.1074/jbc.M101744200. View

13.

Strack S, McNeill R, Colbran R . Mechanism and regulation of calcium/calmodulin-dependent protein kinase II targeting to the NR2B subunit of the N-methyl-D-aspartate receptor. J Biol Chem. 2000; 275(31):23798-806. DOI: 10.1074/jbc.M001471200. View

14.

Thaler C, Koushik S, Puhl 3rd H, Blank P, Vogel S . Structural rearrangement of CaMKIIalpha catalytic domains encodes activation. Proc Natl Acad Sci U S A. 2009; 106(15):6369-74. PMC: 2669344. DOI: 10.1073/pnas.0901913106. View

15.

Borgesius N, van Woerden G, Buitendijk G, Keijzer N, Jaarsma D, Hoogenraad C . βCaMKII plays a nonenzymatic role in hippocampal synaptic plasticity and learning by targeting αCaMKII to synapses. J Neurosci. 2011; 31(28):10141-8. PMC: 6623068. DOI: 10.1523/JNEUROSCI.5105-10.2011. View

16.

Hoelz A, Nairn A, Kuriyan J . Crystal structure of a tetradecameric assembly of the association domain of Ca2+/calmodulin-dependent kinase II. Mol Cell. 2003; 11(5):1241-51. DOI: 10.1016/s1097-2765(03)00171-0. View

17.

Coultrap S, Buard I, Kulbe J, DellAcqua M, Bayer K . CaMKII autonomy is substrate-dependent and further stimulated by Ca2+/calmodulin. J Biol Chem. 2010; 285(23):17930-7. PMC: 2878555. DOI: 10.1074/jbc.M109.069351. View

18.

Pettersen E, Goddard T, Huang C, Couch G, Greenblatt D, Meng E . UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem. 2004; 25(13):1605-12. DOI: 10.1002/jcc.20084. View

19.

Shibata A, Maebashi H, Nakahata Y, Nabekura J, Murakoshi H . Development of a molecularly evolved, highly sensitive CaMKII FRET sensor with improved expression pattern. PLoS One. 2015; 10(3):e0121109. PMC: 4370617. DOI: 10.1371/journal.pone.0121109. View

20.

Kolodziej S, Hudmon A, Waxham M, STOOPS J . Three-dimensional reconstructions of calcium/calmodulin-dependent (CaM) kinase IIalpha and truncated CaM kinase IIalpha reveal a unique organization for its structural core and functional domains. J Biol Chem. 2000; 275(19):14354-9. DOI: 10.1074/jbc.275.19.14354. View