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Published on: July 30, 2014
The effect of toxofilin on the structure and dynamics of monomeric actin
Lívia Czimbalek1, Veronika Kollár1, Roland Kardos1
1University of Pécs, Medical School, Department of Biophysics, Pécs, Szigeti Str. 12, H-7624, Hungary.
Abstract:
The effects of toxofilin (an actin binding protein of Toxoplasma gondii) on G-actin was studied with spectroscopy techniques. Fluorescence anisotropy measurements proved that G-actin and toxofilin interact with 2:1 stoichiometry. The affinity of toxofilin to actin was also determined with a fluorescence anisotropy assay. Fluorescence quenching experiments showed that the accessibility of the actin bound ε-ATP decreased in the presence of toxofilin. The results can be explained by the shift of the nucleotide binding cleft into a closed conformational state. Differential scanning calorimetry measurements revealed that actin monomers become thermodynamically more stable due to the binding of toxofilin.
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