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Cited by 61Protein Kinase Regulation and GTPase SignalingProtein Structure and DynamicsSignaling Pathways in Disease

CaMKII binds both substrates and activators at the active site

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Özden, Can, Sloutsky, Roman, Mitsugi, Tomohiro, Santos, Nicholas, Agnello, Emily, Gaubitz, Christl, Foster, Joshua, Lapinskas, Emily +6 more·2022·Cell Reports
DOI 10.1016/j.celrep.2022.111064PMC PMC9336311

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Abstract

ABSTRACT Ca 2+ /calmodulin dependent protein kinase II (CaMKII) is a signaling protein required for long-term memory. Once activated by Ca 2+ /CaM, it sustains activity even after the Ca 2+ dissipates. In addition to well-known autophosphorylation-mediated mechanism, interaction with specific binding partners also persistently activates CaMKII. A longstanding model invokes two distinct S- and T-sites. If an interactor binds at the T-site, it will preclude autoinhibition and allow substrates to be phosphorylated at the S-site. Here, we specifically test this model with X-ray crystallography, molecular dynamics simulations, and biochemistry. Our data are inconsistent with this model. Co-crystal structures of four different activators or substrates show that they all bind to a single continuous site across the kinase domain. We propose a mechanistic model that persistent CaMKII activity is facilitated by high affinity binding partners, which kinetically compete with autoinhibition by the regulatory segment to allow substrate phosphorylation.

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  • Ca2+/CaM binding peels off the regulatory segment from the kinase surface, unlocking catalytic competency and providing access to substrates and interacting proteins.

    “The binding of Ca2+/CaM peels off the regulatory segment from the surface of the kinase, thus unlocking constraints on catalytic competency and providing access to substrates and CaMKII interacting and anchoring proteins (e.g., GluN2B, Tiam1, and CaMKIIN) (61).”

    From Synaptic memory and CaMKII (2023) · cites this paper as [61]

  • The GluN2B CTD peptide binds in place of the regulatory domain across the entire groove on the kinase, including the active site.

    “As proposed, the CTD peptide does bind in place of the regulatory domain, across the entire groove on the kinase, including the active site (61).”

    From Synaptic memory and CaMKII (2023) · cites this paper as [61]

  • CaMKIINtides inhibit kinase activity with high affinity by binding across the entire substrate binding site and therefore also block GluN2B binding.

    “They inhibit kinase activity with high affinity by binding across the entire substrate binding site (FIGURE 2B7) (61, 147, 153), and therefore also block GluN2B binding (152, 154).”

    From Synaptic memory and CaMKII (2023) · cites this paper as [61]

  • The primary interaction in the CaMKII-GluN2B complex is between the CTD (C-tail) of the GluN2B subunit and the surface groove in the kinase domain of CaMKII exposed by its activation.

    “The primary interaction in the CaMKII-GluN2B complex is between the CTD, also referred to as C-tail, of the GluN2B subunit and the surface groove in the kinase domain of CaMKII exposed by its activation (61).”

    From Synaptic memory and CaMKII (2023) · cites this paper as [61]

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Cited by

Sorted by citations

Authors
Özden, C., Sloutsky, R., Mitsugi, T., Santos, N., Agnello, E., Gaubitz, C., Foster, J., Lapinskas, E., Esposito, E. A., Saneyoshi, T., Kelch, B. A., Garman, S. C., Hayashi, Y., & Stratton, M. M.
Affiliations
  • University of Massachusetts Amherst (US)
  • Kyoto University (JP)
  • University of Massachusetts Chan Medical School (US)
Corresponding author
Stratton, Margaret M.
ORCID
Venue
Cell Reports
Publisher
Elsevier BV
Published
2022-07
Volume
40
Issue
2
Pages
111064
ISSN
2211-1247 (ISSN-L)
DOI
10.1016/j.celrep.2022.111064
PMC
PMC9336311
OpenAlex
W4285097371
Cited by
61
References
8
Citations by year
2026: 19 · 2025: 15 · 2024: 17 · 2023: 7 · 2022: 2 · 2021: 1
FWCI
3.35
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Reports (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion, cc-by-nc-nd)
Funders
  • Ministry of Education, Culture, Sports, Science and Technology (JP18H05434)
  • Japan Science and Technology Agency (CREST JPMJCR20E4)
  • Fujitsu
  • National Institute of General Medical Sciences (R01123157)
  • Kyoto University
  • Japan Society for the Promotion of Science (JP18H04733)
  • Human Frontier Science Program (RGP0020/2019)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
Keywords
CaMKII, calcium/calmodulin-dependent protein kinase II, kinase domain, substrate binding, autoinhibition, autophosphorylation, co-crystal structure, long-term memory, protein-protein interaction
MeSH terms
Calcium, Phosphorylation, Protein Processing, Post-Translational, Catalytic Domain, Calcium-Calmodulin-Dependent Protein Kinase Type 2

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