Abstract
Proteins have numerous physiological functions in the body. Their most important property is serving as depot carriers and as target molecules for many drugs. Three-dimensional fluorescence spectroscopy, zeta potential analysis, resonance light scattering (RLS), thermodynamic analysis, circular dichroism and fluorescence resonance energy transfer (FRET) measurements were performed to study the interactions between estradiol and human lactoferrin (Hlf). According to the three-dimensional fluorescence spectra, the binding of the drug to Hlf induced a slight destabilization of the protein. Circular dichroism results provided new insight into the course of structural transitions of Hlf after addition of estradiol, and revealed that a molten globule state was intermediate between the native and unfolded states. This is a compact state with a significant amount of native-like secondary structure, but a largely disordered tertiary structure. The experimental results further indicated that the fluorescence quenching mechanism between Hlf and estradiol corresponded to a static quenching procedure. The binding constant and the number of binding sites were obtained from modified Stern–Volmer diagrams. The distance r between donor and acceptor was obtained by FRET and found to be 1.96 nm for estradiol and Hlf. The critical induced aggregation concentrations (CCIAC) of estradiol on Hlf were determined by a non-linear relationship between the enhancement of the RLS intensities and drug concentrations. The zeta potential results were used to verify the existence of complexes and confirmed the CCIAC values obtained by the RLS technique. In addition, molecular modeling showed that estradiol was bound within the domain II of the C-lobe and N-lobe of the Hlf via hydrophobic interactions and hydrogen bonding.
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The financial support of the Research Council of the Mashhad Branch, Islamic Azad University is gratefully acknowledged. The authors thank Dr. Ljungberg for the English editing.
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Moastajabodave, Z., Beigoli, S., Sharifi Rad, A. et al. Structural transition of lactoferrin upon interaction with estradiol as revealed by spectroscopic techniques: a molten globule state investigation. J IRAN CHEM SOC 15, 2159–2173 (2018). https://doi.org/10.1007/s13738-018-1409-9
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DOI: https://doi.org/10.1007/s13738-018-1409-9