Biophysical study of phloretin with human serum albumin in liposomes using spectroscopic methods
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ORIGINAL ARTICLE
Biophysical study of phloretin with human serum albumin in liposomes using spectroscopic methods Seda Karabulut1 · Mahmut Toprak1 Received: 4 February 2020 / Revised: 3 July 2020 / Accepted: 19 July 2020 © European Biophysical Societies’ Association 2020
Abstract The ability of drugs to diffuse through the lipid bilayer of cell membranes is important for their metabolism, distribution, and efficacy. In this study, the interaction between phloretin and human serum albumin (HSA) in an L-egg lecithin phosphatidylcholine (PC) liposome suspension was investigated by fluorescence and absorbance spectroscopy. The spectroscopic and fluorescence quenching experiments show that phloretin molecules penetrated into the lumen of the liposome. The partition coefficient of phloretin in the PC liposome suspensions was calculated from fluorescence quenching measurements. The results show that phloretin efficiently quenches the intrinsic fluorescence of HSA through a combination of dynamic and static quenching. The values of Gibbs free energy, and the enthalpy and entropic change in the binding process of phloretin with HSA in the PC liposome suspensions were negative, suggesting that the binding process of phloretin and HSA was spontaneous. Hydrogen bonding and van der Waals force interactions play an important role in the interaction between the two molecules. In addition, binding of phloretin to HSA in liposome suspensions was investigated by synchronous fluorescence spectroscopy. Keywords Phloretin · HSA · Static quenching · Dynamic quenching · Phosphatidylcholine Abbreviations HSA Human serum albumin PC L-egg lecithin phosphatidylcholine
Introduction Liposomes are made of polar lipids that form bilayers encompassing an aqueous compartment and are suitable models of cell membranes considering the structural similarity with biological membranes (Rodrigues et al. 2001; Evans et al. 2015; Strohmeier et al. 2016). Liposomes are successfully applied in studies of interactions of phospholipids and drugs at a cellular level as they are more suitable systems for determination of partition coefficients than the conventional octanol/buffer system (Eyer et al. 2014; Neunert et al. 2015). The partition coefficient expresses compound distribution between the lipid and water phases. This distribution is connected with lipophilicity, charge, size, and hydrogen-bonding properties * Mahmut Toprak [email protected]; [email protected] 1
Department of Chemistry, Bingol University, 12000 Bingol, Turkey
of compounds, which affects the extent of their permeation into the cell membranes (Przybylo et al. 2014; Takegami et al. 2015; Toprak 2016). The kinetics of lipid bilayer permeations have received extensive attention in the field of drug development and toxicology (Ikonen et al. 2007; Trusova et al. 2013). The partition coefficient has been attributed to evaluate its pharmacokinetics and toxicity kinetic properties that include absorption, distribution, metabolism, toxicity, and excretion (Auner et al. 2005; Bao et
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