title: Two-Dimensional Potentials of Mean Force of Nile Red in Intact and Damaged Model Bilayers. Application to Calculations of Fluorescence Spectra.
abstract: Fluorescent dyes revolutionized and expanded our understanding of biological membranes. The interpretation of experimental fluorescence data in terms of membrane structure, however, requires detailed information about the molecular environment of the dyes. Nile red is a fluorescent molecule whose excitation and emission maxima depend on the polarity of the solvent. It is mainly used as a probe to study lipid microenvironments, for example in imaging the progression of damage to the myelin sheath in multiple sclerosis. In this study, we determine the position and orientation of Nile red in lipid bilayers by calculating two-dimensional Potential of Mean Force (2D-PMF) profiles in a defect-free 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer and in damaged bilayers containing two mixtures of the oxidized lipid 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine and POPC. From 2D-PMF simulations we obtain positions and orientations of Nile Red corresponding to the minimum on the binding free energy surface in three different membrane environments with increasing amounts of water, mimicking damage in biological tissue. Using representative snapshots from the simulations, we use combined quantum mechanical/molecular mechanical (QM/MM) models to calculate the emission spectrum of Nile red as a function of its local solvation environment. The results of QM and QM/MM computations are in qualitative agreement with the experimentally observed shift in fluorescence for the dye moving from aqueous solution to the more hydrophobic environment of the lipid interiors. The range of the conformation dependent values of the computed absorption-emission spectra and the lack of solvent relaxation effects in the QM/MM calculations made it challenging to delineate specific differences between the intact and damaged bilayers.
authors.affiliation: Department of Biological Sciences and Centre for Molecular Simulation, University of Calgary , 2500 University Drive N.W., Calgary, Alberta T2N1N4, Canada.
authors.forename: Gurpreet
authors.initials: G
authors.lastname: Singh
authors.affiliation: Department of Biological Sciences and Centre for Molecular Simulation, University of Calgary , 2500 University Drive N.W., Calgary, Alberta T2N1N4, Canada.
authors.forename: Adam C
authors.initials: AC
authors.lastname: Chamberlin
authors.affiliation: Department of Biological Sciences and Centre for Molecular Simulation, University of Calgary , 2500 University Drive N.W., Calgary, Alberta T2N1N4, Canada.
authors.forename: Hristina R
authors.initials: HR
authors.lastname: Zhekova
authors.affiliation: Department of Biological Sciences and Centre for Molecular Simulation, University of Calgary , 2500 University Drive N.W., Calgary, Alberta T2N1N4, Canada.
authors.forename: Sergei Y
authors.initials: SY
authors.lastname: Noskov
authors.affiliation: Department of Biological Sciences and Centre for Molecular Simulation, University of Calgary , 2500 University Drive N.W., Calgary, Alberta T2N1N4, Canada.
authors.forename: D Peter
authors.initials: DP
authors.lastname: Tieleman
mesh_terms: D000465:Algorithms; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D010078:Oxazines / Q000737:chemistry* / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry; D011789:Quantum Theory; D013050:Spectrometry, Fluorescence
chemical_list: D008051:Lipid Bilayers; D010078:Oxazines; D010713:Phosphatidylcholines; C044808:nile red; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine