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Isolation of a CK2α Subunit and the Holoenzyme from the Mussel Mytilus galloprovincialis and Construction of the CK2α and CK2β cDNAs

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Abstract

Protein kinase CK2 is a ubiquitous, highly pleiotropic, and constitutively active phosphotransferase that phosphorylates mainly serine and threonine residues. CK2 has been studied and characterized in many organisms, from yeast to mammals. The holoenzyme is generally composed of two catalytic (α and/or α′) and two regulatory (β) subunits, forming a differently assembled tetramer. The free and catalytically active α/α′ subunits can be present in cells under some circumstances. We present here the isolation of a putative catalytic CK2α subunit and holoenzyme from gills of the mussel Mytilus galloprovincialis capable of phosphorylating the purified recombinant ribosomal protein rMgP1. For further analysis of M. galloprovincialis protein kinase CK2, the cDNA molecules of CK2α and CK2β subunits were constructed and cloned into expression vectors, and the recombinant proteins were purified after expression in Escherichia coli. The recombinant MgCK2β subunit and MgP1 were phosphorylated by the purified recombinant MgCK2α subunit. The mussel enzyme presented features typical for CK2: affinity for GTP, inhibition by both heparin and ATP competitive inhibitors (TBBt, TBBz), and sensitivity towards NaCl. Predicted amino acid sequence comparison showed that the M. galloprovincialis MgCK2α and MgCK2β subunits have similar features to their mammalian orthologs.

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Acknowledgements

The authors are grateful to Dr. Y. Drosos, Miss A. Sielouvari, and Dr. K. Kubiński for scientific help and discussion. 4,5,6,7-Tetrabromo-1H-benzotriazole (TBBt) and 4,5,6,7-tetrabromo-1H-benzimidazole (TBBz) were generous gifts of Zygmunt Kazimierczuk (Department of Experimental Pharmacology, Polish Academy of Sciences, Medical Research Center, Warsaw, Poland). This work was supported by the University of Athens, the Special Account for Research Grants of Athens University (SARG, 70/4/7803 to SK), and the Greek National Fellowship Foundation (IKY) (fellowship to RMK).

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens, 15701, Greece

    Regina-Maria Kolaiti & Sophia Kouyanou-Koutsoukou

  2. Department of Molecular Biology, Institute of Biotechnology, The John Paul II Catholic University of Lublin, Al. Krasnicka 102, 20-718, Lublin, Poland

    Andrea Baier & Ryszard Szyszka

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  1. Regina-Maria Kolaiti
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  2. Andrea Baier
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  3. Ryszard Szyszka
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  4. Sophia Kouyanou-Koutsoukou
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Correspondence to Sophia Kouyanou-Koutsoukou.

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R.-M. Kolaiti and A. Baier gave equal contribution to this study.

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Kolaiti, RM., Baier, A., Szyszka, R. et al. Isolation of a CK2α Subunit and the Holoenzyme from the Mussel Mytilus galloprovincialis and Construction of the CK2α and CK2β cDNAs. Mar Biotechnol 13, 505–516 (2011). https://doi.org/10.1007/s10126-010-9321-z

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