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Thermal and dynamic behaviour of the actin monomer in case of different cations

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Abstract

Actin is one of the important elements of the striated muscle that transmits force from the myosin filaments and as a part of the cytoskeleton plays an important role in shape determination of cells. It is a known experience that removal of the divalent cation affects the dynamic behaviour of actin in both forms. Paramagnetic probes and electron paramagnetic resonance (EPR) spectroscopy provide direct technique by which the rotation and the orientation of specifically labelled proteins can be followed during biochemical manipulations. The spectroscopic measurements could be combined with DSC measurements that report domain stability and interactions and allow the calculation of the thermodynamic parameters during the melting process. Actin was spin-labelled with maleimide or fluoro-dinitro proxyl probe molecules which are bound to the Cys-374 or Lys-61 residues of the smaller domain. EPR spectroscopy spectra were recorded in monomer form in Ca- and EGTA-state as a function of temperature up to the melting point. Similarly, DSC measurements were performed and analyzed using the kinetic theory. The measurements showed that removal of the divalent cation from the globular actin induced significant local and global structural change both in the thermodynamic properties and the rotational mobility of actin detected by DSC and EPR. On the basis of the results derived by deconvolution of the DSC pattern we can suggest a non-interactive two-domain melting for the monomer actin after removing the divalent cations.

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Acknowledgements

The SETARAM Micro DSC-II was purchased with a grant (CO-272) from the Hungarian Scientific Research Fund (Dénes Lőrinczy). ESP 300E EPR spectrometer was purchased with a grant (CO-123) from the Hungarian Scientific Research Fund (Joseph Belagyi).

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Authors and Affiliations

  1. Institute of Forensic Medicine, University of Pécs, Szigeti str. 12, Pecs, 7624, Hungary

    F. Könczöl

  2. Institute of Biophysics, Faculty of Medicine, University of Pécs, Szigeti str. 12, Pecs, 7624, Hungary

    J. Belagyi & D. Lőrinczy

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  1. F. Könczöl
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  2. J. Belagyi
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  3. D. Lőrinczy
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Correspondence to D. Lőrinczy.

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Könczöl, F., Belagyi, J. & Lőrinczy, D. Thermal and dynamic behaviour of the actin monomer in case of different cations. J Therm Anal Calorim 111, 1717–1723 (2013). https://doi.org/10.1007/s10973-012-2206-9

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  • DOI: https://doi.org/10.1007/s10973-012-2206-9

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