Interaction between CK2α and CK2β, the Subunits of Protein Kinase CK2: Thermodynamic Contributions of Key Residues on the CK2α Surface
| العنوان: | Interaction between CK2α and CK2β, the Subunits of Protein Kinase CK2: Thermodynamic Contributions of Key Residues on the CK2α Surface |
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| المؤلفون: | David W. Litchfield, J. Raaf, Karsten Klopffleisch, Birgitte Brinkmann Olsen, Greg Vilk, Elena Brunstein, Olaf-Georg Issinger, Nils Bischoff, Karsten Niefind |
| المصدر: | Biochemistry Publications Raaf, J, Bischoff, N, Kloppfleisch, K, Brunstein, E, Olsen, B B, Vilk, G, Litchfield, D, Issinger, O-G & Niefind, K 2011, ' Interaction between CK2α and CK2β, the Subunits of Protein Kinase CK2 : Thermodynamic Contributions of Key Residues on the CK2α Surface ', Biochemistry, vol. 50, no. 4, pp. 512-22 . https://doi.org/10.1021/bi1013563 |
| بيانات النشر: | Scholarship@Western, 2011. |
| سنة النشر: | 2011 |
| مصطلحات موضوعية: | animal structures, Dimer, Phenylalanine, Isozyme, Biochemistry, chemistry.chemical_compound, Leucine, Humans, Casein Kinase II, Alanine, Kinase, Chemistry, fungi, Temperature, Membrane Proteins, Hydrogen-Ion Concentration, Peptide Fragments, Isoenzymes, Membrane protein, Amino Acid Substitution, Thermodynamics, Casein kinase 2 |
| الوصف: | The protein Ser/Thr kinase CK2 (former name: casein kinase II) exists predominantly as a heterotetrameric holoenzyme composed of two catalytic subunits (CK2α) bound to a dimer of noncatalytic subunits (CK2β). We undertook a study to further understand how these subunits interact to form the tetramer. To this end, we used recombinant, C-terminal truncated forms of human CK2 subunits that are able to form the holoenzyme. We analyzed the interaction thermodynamics between the binding of CK2α and CK2β as well as the impact of changes in temperature, pH, and the ionization enthalpy of the buffer using isothermal titration calorimetry (ITC). With structure-guided alanine scanning mutagenesis we truncated individual side chains in the hydrophobic amino acid cluster located within the CK2α interface to identify experimentally the amino acids that dominate affinity. The ITC results indicate that Leu41 or Phe54 single mutations were most disruptive to binding of CK2β. Additionally, these CK2α mutants retained their kinase activity. Furthermore, the substitution of Leu41 in combination with Phe54 showed that the individual mutations were not additive, suggesting that the cooperative action of both residues played a role. Interestingly, the replacement of Ile69, which has a central position in the interaction surface of CK2α, only had modest effects. The differences between Leu41, Phe54, and Ile69 in interaction relevance correlate with solvent accessibility changes during the transition from unbound to CK2β-bound CK2α. Identifying residues on CK2α that play a key role in CK2α/CK2β interactions is important for the future generation of small molecule drug design. |
| DOI: | 10.1021/bi1013563 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::89f85a860fecb9748e1585cdcf1c5a9b https://ir.lib.uwo.ca/biochempub/144 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....89f85a860fecb9748e1585cdcf1c5a9b |
| قاعدة البيانات: | OpenAIRE |
| DOI: | 10.1021/bi1013563 |
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