The Protein Journal

, Volume 33, Issue 4, pp 386–393 | Cite as

Thermodynamic Aspects of cAMP Dependent Protein Kinase Catalytic Subunit Allostery

  • Rait Kivi
  • Per Jemth
  • Jaak Järv


Kinetics of thermal inactivation of acrylodan-labeled cAMP dependent protein kinase catalytic subunit, its binary complexes with ATP and peptide inhibitor PKI[5–24], respectively, and the ternary complex involving both of these ligands were studied at different temperatures (5–50 °C). The thermodynamic parameters ΔH and ΔS for ligand binding equilibria as well as for the allosteric interaction between the binding sites of these ligands were obtained by using the Van’t Hoff analysis. The results indicated that more inter- and intra-molecular non-covalent bonds were involved in ATP binding with the protein when compared to the peptide binding. Similarly, nucleotide and peptide binding steps were accompanied with different entropy effects, while almost no entropy change accompanied PKI[5–24] binding, suggesting that the protein flexibility was not affected in this case. Differently from the binary complex formation the ternary complex formation was accompanied by a significant entropy change and with intensive formation of new non-covalent interactions (ΔH). At the same time both ligand binding steps as well as the allosteric interaction between ligand binding sites could be described by a common entropy–enthalpy compensation plot, pointing to a similar mechanism of these phenomena. It was concluded that numerous weak interactions govern the allostery of cAMP dependent protein kinase catalytic subunit.


cAMP-Dependent protein kinase catalytic subunit Protein denaturation Fluorescence spectroscopy Acrylodan Ligand binding Protein stabilization Thermodynamics Allostery 



Acrylodan, 6-acryloyl-2-dimethylaminonaphthalene


Adenosine triphosphate


3-(N-morpholino)propanesulfonic acid


cAMP-Dependent protein kinase A catalytic subunit from Mus musculus





This work was financially supported by the Estonian Ministry of Education and Research, Grants SF0180064s08 and IUT14-20, by Graduate School “Functional materials and technologies” receiving funding from the European Social Fund under Project 1.2.0401.09-0079 in the University of Tartu, and Kristjan Jaak and DoRa T6 Travel Grants to RK.


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Institute of ChemistryUniversity of TartuTartuEstonia
  2. 2.Department of Medical Biochemistry and MicrobiologyUppsala UniversityUppsalaSweden

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