Abstract
Phloretin is a known modifier of the internal dipole potential of lipid membranes. We studied the interaction of phloretin with model lipid membranes and how it influences the membrane dipole organization using ANS as fluorescent probe. The fluorescence increase observed when ANS binds to DMPC liposomes in gel phase (13 °C) was 2.5 times larger in the presence of phloretin. This effect was due to an increase in ANS affinity, which can be related to the known capability of phloretin in decreasing the dipole potential. Conversely, when the experiments were carried out at 33 °C (liquid crystalline phase), phloretin completely inhibited the increase in ANS fluorescence. In addition, phloretin only affected the electrical properties of the membrane in the gel phase, whereas it modifies structural ones in the liquid-crystalline state. We postulate that phloretin was bound only to the DMPC interface in the gel phase decreasing the surface negative charge density without modifying the structural properties of the ANS binding sites. In the liquid-crystalline phase instead, it increased the accessibility of water to the ANS binding sites decreasing the intrinsic affinity and the fluorescence quantum yield of ANS.
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Acknowledgments
This work has been carried out with grants from CONICET (PIP 0307) and ANPCyT (PICT 2005/38056) to O.A.R. and also with support from the Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario. O.A.R. and G.M. are Members of the Carrera del Investigador from CONICET. A.C.C. was Fellow of ANPCyT (FONCyT) and of CONICET. The authors would like to thank E.A. Disalvo for the critical reading of this manuscript.
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Cutró, A.C., Montich, G. & Roveri, O.A. Effect of Phloretin on the Binding of 1-Anilino-8-naphtalene sulfonate (ANS) to 1,2-Dimyristoyl-sn-glycero-3-phosphocoline (DMPC) Vesicles in the Gel and Liquid-Crystalline State. J Membrane Biol 248, 137–144 (2015). https://doi.org/10.1007/s00232-014-9750-0
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DOI: https://doi.org/10.1007/s00232-014-9750-0