Insights into the regulation of eukaryotic elongation factor 2 kinase and the interplay between its domains
| Autor: | Christopher G. Proud, Stephen J. Finn, Claire E. Moore, Craig R. Pigott, Halina Mikolajek, Curtis W. Phippen, Jörn M. Werner |
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| Rok vydání: | 2011 |
| Předmět: |
Biochemistry
SH3 domain MAP2K7 mTORC1 mammalian target of rapamycin complex 1 0302 clinical medicine Catalytic Domain GST glutathione transferase ATP-γS adenosine 5′-[γ-thio]triphosphate Phosphorylation 0303 health sciences education.field_of_study CaM calmodulin biology HRP horseradish peroxidase SEL1 domain Ni-NTA Ni2+-nitrilotriacetic acid Cell biology Zinc MHCKA myosin heavy-chain kinase A LB Luria–Bertani α-kinase eEF2K eEF2 kinase Research Article TRPM transient receptor potential melastatin-like Elongation Factor 2 Kinase STD saturation transfer difference EEF2 TEV tobacco etch virus 03 medical and health sciences HEK human embryonic kidney Humans c-Raf Amino Acid Sequence eEF2 eukaryotic elongation factor 2 education Protein kinase A Molecular Biology Nuclear Magnetic Resonance Biomolecular 030304 developmental biology Cyclin-dependent kinase 1 Binding Sites Sequence Homology Amino Acid Cyclin-dependent kinase 2 Cell Biology Protein Structure Tertiary HEK293 Cells biology.protein Elongation Factor-2 Kinase calmodulin (CaM) eukaryotic elongation factor 2 (eEF2) 030217 neurology & neurosurgery |
| Zdroj: | Biochemical Journal |
| ISSN: | 1470-8728 |
| Popis: | eEF2K (eukaryotic elongation factor 2 kinase) is a Ca2+/CaM (calmodulin)-dependent protein kinase which regulates the translation elongation machinery. eEF2K belongs to the small group of so-called ‘α-kinases’ which are distinct from the main eukaryotic protein kinase superfamily. In addition to the α-kinase catalytic domain, other domains have been identified in eEF2K: a CaM-binding region, N-terminal to the kinase domain; a C-terminal region containing several predicted α-helices (resembling SEL1 domains); and a probably rather unstructured ‘linker’ region connecting them. In the present paper, we demonstrate: (i) that several highly conserved residues, implicated in binding ATP or metal ions, are critical for eEF2K activity; (ii) that Ca2+/CaM enhance the ability of eEF2K to bind to ATP, providing the first insight into the allosteric control of eEF2K; (iii) that the CaM-binding/α-kinase domain of eEF2K itself possesses autokinase activity, but is unable to phosphorylate substrates in trans; (iv) that phosphorylation of these substrates requires the SEL1-like domains of eEF2K; and (v) that highly conserved residues in the C-terminal tip of eEF2K are essential for the phosphorylation of eEF2, but not a peptide substrate. On the basis of these findings, we propose a model for the functional organization and control of eEF2K. |
| Databáze: | OpenAIRE |
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