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14-3-3 protein directly interacts with the kinase domain of calcium/calmodulin-dependent protein kinase kinase (CaMKK2).

Psenakova K, Petrvalska O, Kylarova S, Lentini Santo D, Kalabova D, Herman P, Obsilova V, Obsil T
Biochim Biophys Acta Gen Subj 1862 1612-1625 (2018)
Cited: 13 times
EuropePMC logo PMID: 29649512

Abstract

Background

Calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is a member of the Ca2+/calmodulin-dependent kinase (CaMK) family involved in adiposity regulation, glucose homeostasis and cancer. This upstream activator of CaMKI, CaMKIV and AMP-activated protein kinase is inhibited by phosphorylation, which also triggers an association with the scaffolding protein 14-3-3. However, the role of 14-3-3 in the regulation of CaMKK2 remains unknown.

Methods

The interaction between phosphorylated CaMKK2 and the 14-3-3γ protein, as well as the architecture of their complex, were studied using enzyme activity measurements, small-angle x-ray scattering (SAXS), time-resolved fluorescence spectroscopy and protein crystallography.

Results

Our data suggest that the 14-3-3 protein binding does not inhibit the catalytic activity of phosphorylated CaMKK2 but rather slows down its dephosphorylation. Structural analysis indicated that the complex is flexible and that CaMKK2 is located outside the phosphopeptide-binding central channel of the 14-3-3γ dimer. Furthermore, 14-3-3γ appears to interact with and affect the structure of several regions of CaMKK2 outside the 14-3-3 binding motifs. In addition, the structural basis of interactions between 14-3-3 and the 14-3-3 binding motifs of CaMKK2 were elucidated by determining the crystal structures of phosphopeptides containing these motifs bound to 14-3-3.

Conclusion

14-3-3γ protein directly interacts with the kinase domain of CaMKK2 and the region containing the inhibitory phosphorylation site Thr145 within the N-terminal extension.

Reviews - 6eww mentioned but not cited (1)

  1. The 14-3-3 Proteins as Important Allosteric Regulators of Protein Kinases. Obsilova V, Obsil T. Int J Mol Sci 21 E8824 (2020)


Reviews citing this publication (5)

  1. Molecular Mechanisms Underlying Ca2+/Calmodulin-Dependent Protein Kinase Kinase Signal Transduction. Tokumitsu H, Sakagami H. Int J Mol Sci 23 11025 (2022)
  2. Structural insights into the functional roles of 14-3-3 proteins. Obsilova V, Obsil T. Front Mol Biosci 9 1016071 (2022)
  3. CaMKK2 as an emerging treatment target for bipolar disorder. Kaiser J, Nay K, Horne CR, McAloon LM, Fuller OK, Muller AG, Whyte DG, Means AR, Walder K, Berk M, Hannan AJ, Murphy JM, Febbraio MA, Gundlach AL, Scott JW. Mol Psychiatry (2023)
  4. Challenges of studying 14-3-3 protein-protein interactions with full-length protein partners. Somsen BA, Ottmann C. Biophys J 121 1115-1116 (2022)
  5. Modulating GPCR and 14-3-3 protein interactions: Prospects for CNS drug discovery. Kongsamut S, Eishingdrelo H. Drug Discov Today 28 103641 (2023)

Articles citing this publication (7)

  1. CaMKK2 is inactivated by cAMP-PKA signaling and 14-3-3 adaptor proteins. Langendorf CG, O'Brien MT, Ngoei KRW, McAloon LM, Dhagat U, Hoque A, Ling NXY, Dite TA, Galic S, Loh K, Parker MW, Oakhill JS, Kemp BE, Scott JW. J Biol Chem 295 16239-16250 (2020)
  2. 14-3-3 proteins inactivate DAPK2 by promoting its dimerization and protecting key regulatory phosphosites. Horvath M, Petrvalska O, Herman P, Obsilova V, Obsil T. Commun Biol 4 986 (2021)
  3. Set-up and screening of a fragment library targeting the 14-3-3 protein interface. Valenti D, Neves JF, Cantrelle FX, Hristeva S, Lentini Santo D, Obšil T, Hanoulle X, Levy LM, Tzalis D, Landrieu I, Ottmann C. Medchemcomm 10 1796-1802 (2019)
  4. 14-3-3 protein inhibits CaMKK1 by blocking the kinase active site with its last two C-terminal helices. Petrvalska O, Honzejkova K, Koupilova N, Herman P, Obsilova V, Obsil T. Protein Sci 32 e4805 (2023)
  5. A new soaking procedure for X-ray crystallographic structural determination of protein-peptide complexes. Ballone A, Lau RA, Zweipfenning FPA, Ottmann C. Acta Crystallogr F Struct Biol Commun 76 501-507 (2020)
  6. Interaction of an IκBα Peptide with 14-3-3. Wolter M, Santo DL, Herman P, Ballone A, Centorrino F, Obsil T, Ottmann C. ACS Omega 5 5380-5388 (2020)
  7. Structural insights into regulation of the PEAK3 pseudokinase scaffold by 14-3-3. Torosyan H, Paul MD, Forget A, Lo M, Diwanji D, Pawłowski K, Krogan NJ, Jura N, Verba KA. Nat Commun 14 3543 (2023)


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