Enzymatic Activity of CaMKII Is Not Required for Its Interaction with the Glutamate Receptor Subunit GluN2B

Barcomb, Kelsey; Coultrap, Steven J.; Bayer, Karl

doi:10.1124/mol.113.089045

Cited by 21 publications

(38 citation statements)

References 60 publications

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“…Under basal conditions, CaMKII was present throughout the dendritic shaft and slightly enriched in dendritic spines (Figure 5A). While chemical LTP-stimuli with glutamate/glycine induce robust further CaMKII accumulation in dendritic spines (Barcomb et al, 2013; Bayer et al, 2001; 2006; O'Leary et al, 2011), no such accumulation was seen after chemical LTD-stimuli (Figure 5A; Supplemental Movie 1). However, inhibiting either DAPK1 (with 10 µM TC) or CaN (with 10 µM CycA) allowed significant translocation of endogenous CaMKII into spines even after LTD-stimuli (Figure 5A; Supplemental Movies 2–3), as expected based on our hypothesis.…”

Section: Resultsmentioning

confidence: 99%

“…Cell culture, transfections, protein preparations, and Western analysis were done as previously described (Barcomb et al, 2013; 2014; 2015; Bayer et al, 2001; 2006; Coultrap and Bayer, 2012b; 2014; Goodell et al, 2016; 2014; O'Leary et al, 2011; Singla et al, 2001; Vest et al, 2007) and are detailed in the Supplemental Experimental Procedures. Antibodies and dilutions are shown in Supplemental Table 1.…”

Section: Methodsmentioning

confidence: 99%

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DAPK1 Mediates LTD by Making CaMKII/GluN2B Binding LTP Specific

et al. 2017

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The death associated protein kinase 1 (DAPK1) is a potent mediator of neuronal cell death. Here, we find that DAPK1 also functions in synaptic plasticity by regulating the Ca2+/calmodulin (CaM)-dependent protein kinase II (CaMKII). CaMKII and T286-autophosphorylation are required for both long-term potentiation (LTP) and depression (LTD), two opposing forms of synaptic plasticity underlying learning, memory and cognition. T286-autophosphorylation induces CaMKII binding to the NMDA receptor (NMDAR) subunit GluN2B, which mediates CaMKII synaptic accumulation during LTP. We find that the LTP-specificity of CaMKII synaptic accumulation is due to its LTD-specific suppression by calcineurin (CaN)-dependent DAPK1 activation, which in turn blocks CaMKII binding to GluN2B. This suppression is enabled by competitive DAPK1 versus CaMKII binding to GluN2B. Negative regulation of DAPK1/GluN2B binding by Ca2+/CaM results in synaptic DAPK1 removal during LTP but retention during LTD. A pharmacogenetic approach showed that suppression of CaMKII/GluN2B binding is a DAPK1 function required for LTD.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

DAPK1 Mediates LTD by Making CaMKII/GluN2B Binding LTP Specific

et al. 2017

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“…First, it has been found that nucleotide (ATP or ADP) strongly enhances the ability of CaMKII to bind to NMDAR in the presence of Ca 2+ /CaM (Barcomb et al, 2013; O’Leary et al, 2011; Robison et al, 2005). This can be understood as a nucleotide-dependent stabilization of the binding of pT286 to the catalytic site and the consequent availability of the T site (to which T286 normally binds in the off state) for NMDAR binding.…”

Section: Principle 4: Nmdar Protects Pt286 From Dephosphorylation By mentioning

confidence: 99%

Biochemical principles underlying the stable maintenance of LTP by the CaMKII/NMDAR complex

Lisman¹,

Raghavachari²

2015

Brain Research

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Memory involves the storage of information at synapses by an LTP-like process. This information storage is synapse specific and can endure for years despite the turnover of all synaptic proteins. There must, therefore, be special principles that underlie the stability of LTP. Recent experimental results suggest that LTP is maintained by the complex of CaMKII with the NMDAR. Here we consider the specifics of the CaMKII/NMDAR molecular switch, with the goal of understanding the biochemical principles that underlie stable information storage by synapses. Consideration of a variety of experimental results suggests that multiple principles are involved. One switch requirement is to prevent spontaneous transitions from the off to the on state. The highly cooperative nature of CaMKII autophosphorylation by Ca2+ (Hill coefficient of 8) and the fact that formation of the CaMKII/NMDAR complex requires release of CaMKII from actin are mechanisms that stabilize the off state. The stability of the on state depends critically on intersubunit autophosphorylation, a process that restores any loss of pT286 due to phosphatase activity. Intersubunit autophosphorylation is also important in explaining why on state stability is not compromised by protein turnover. Recent evidence suggests that turnover occurs by subunit exchange. Thus, stability could be achieved if a newly inserted unphosphorylated subunit was autophosphorylated by a neighboring subunit. Based on other recent work, we posit a novel mechanism that enhances the stability of the on state by protection of pT286 from phosphatases. We posit that the binding of the NMNDAR to CaMKII forces pT286 into the catalytic site of a neighboring subunit, thereby protecting pT286 from phosphatases. A final principle concerns the role of structural changes. The binding of CaMKII to the NMDAR may act as a tag to organize the binding of further proteins that produce the synapse enlargement that underlies late LTP. We argue that these structural changes not only enhance transmission, but also enhance the stability of the CaMKII/NMDAR complex. Together, these principles provide a mechanistic framework for understanding how individual synapses produce stable information storage.

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“…However, while GluN2B binding is induced by CaMKII stimulation, actual enzymatic CaMKII activity is not required, neither in vitro nor within cells [13]. This raised the question if other positive-regulatory factors could also directly induce the CaMKII/GluN2B binding even in absence of stimuli that induce CaMKII activity.…”

Section: Discussionmentioning

confidence: 99%

“…For this reason, CaMKII binding to GluN2B is enhanced less by the presence of ATP compared to ADP [12], as ADP cannot be used for the inhibitory phosphorylation reactions. However, due to the positive effect of direct nucleotide binding to CaMKII, ATP still enhances the binding to GluN2B compared to conditions without any nucleotide present [12], [13]. Second, CaMKII autophosphorylation at T305/306 prevents binding of Ca 2+ /CaM to CaMKII [20], [21], and can thereby also inhibit CaMKII binding to GluN2B [12], as Ca 2+ /CaM is required to induced binding to GluN2B, either directly or via autophosphorylation at T286.…”

Section: Introductionmentioning

confidence: 99%

CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents

Goodell¹,

Eliseeva²,

Coultrap³

et al. 2014

PLoS ONE

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Binding of the Ca2+/calmodulin(CaM)-dependent protein kinase II (CaMKII) to the NMDA-type glutamate receptor (NMDAR) subunit GluN2B controls long-term potentiation (LTP), a form of synaptic plasticity thought to underlie learning and memory. Regulation of this interaction is well-studied biochemically, but not under conditions that mimic the macromolecular crowding found within cells. Notably, previous molecular crowding experiments with lysozyme indicated an effect on the CaMKII holoenzyme conformation. Here, we found that the effect of molecular crowding on Ca2+/CaM-induced CaMKII binding to immobilized GluN2B in vitro depended on the specific crowding reagent. While binding was reduced by lysozyme, it was enhanced by BSA. The ATP content in the BSA preparation caused CaMKII autophosphorylation at T286 during the binding reaction; however, enhanced binding was also observed when autophosphorylation was blocked. Importantly, the positive regulation by nucleotide and BSA (as well as other macromolecular crowding reagents) did not alleviate the requirement for CaMKII stimulation to induce GluN2B binding. The differential effect of lysozyme (14 kDa) and BSA (66 kDa) was not due to size difference, as both dextran-10 and dextran-70 enhanced binding. By contrast, crowding with immunoglobulin G (IgG) reduced binding. Notably, lysozyme and IgG but not BSA directly bound to Ca2+/CaM in an overlay assay, suggesting a competition of lysozyme and IgG with the Ca2+/CaM-stimulus that induces CaMKII/GluN2B binding. However, lysozyme negatively regulated binding even when it was instead induced by CaMKII T286 phosphorylation. Alternative modes of competition would be with CaMKII or GluN2B, and the negative effects of lysozyme and IgG indeed also correlated with specific or non-specific binding to the immobilized GluN2B. Thus, the effect of any specific crowding reagent can differ, depending on its additional direct effects on CaMKII/GluN2B binding. However, the results of this study also indicate that, in principle, macromolecular crowding enhances CaMKII binding to GluN2B.

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Enzymatic Activity of CaMKII Is Not Required for Its Interaction with the Glutamate Receptor Subunit GluN2B

Cited by 21 publications

References 60 publications

DAPK1 Mediates LTD by Making CaMKII/GluN2B Binding LTP Specific

DAPK1 Mediates LTD by Making CaMKII/GluN2B Binding LTP Specific

Biochemical principles underlying the stable maintenance of LTP by the CaMKII/NMDAR complex

CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents

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