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Fluorescence of nitrobenzoxadiazole (NBD) labeled lipids in model membranes is connected not to lipid mobility, but to probe location
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SYSNO ASEP 0453494 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Fluorescence of nitrobenzoxadiazole (NBD) labeled lipids in model membranes is connected not to lipid mobility, but to probe location Tvůrce(i) Amaro, Mariana (UFCH-W) RID, ORCID
Filipe, H. A. (PT)
Ramalho, J. P. P. (PT)
Hof, Martin (UFCH-W) RID, ORCID
Loura, L. M. S. (PT)Zdroj.dok. Physical Chemistry Chemical Physics. - : Royal Society of Chemistry - ISSN 1463-9076
Roč. 18, č. 10 (2016), s. 7042-7054Poč.str. 13 s. Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova fluorescence ; nitrobenzoxadiazole Vědní obor RIV CF - Fyzikální chemie a teoretická chemie CEP GBP208/12/G016 GA ČR - Grantová agentura ČR Institucionální podpora UFCH-W - RVO:61388955 UT WOS 000371608600011 EID SCOPUS 84960153667 DOI 10.1039/c5cp05238f Anotace Nitrobenzoxadiazole (NBD)-labeled lipids are popular fluorescent membrane probes. However, understanding of important aspects of NBD photophysics remains incomplete, including the observed shift in the emission spectrum of NBD-lipids to longer wavelengths following excitation at the red-edge of the absorption spectrum (red-edge excitation shift or REES). REES of NBD-lipids has been previously interpreted as reflecting restricted mobility of solvent surrounding the fluorophore in membrane environments. However, this requires a large change in dipole moment (Δμ) of NBD upon excitation. Previous calculations of Δμ of NBD in the literature have been carried out using outdated semiempirical methods, leading to conflicting values. Using up-to-date density functional theory methods, we recalculated Δμ confirming a rather small value (~2 D). Fluorescence measurements confirmed a REES of ~16 nm for 1,2-dioleoyl-sn-glycero-3-phospho-L-serine-N-(NBD) (NBD-PS) in dioleoylphosphatidylcholine vesicles. However, the observed shift is independent on both temperature and presence of cholesterol, and is therefore insensitive to membrane mobility and hydration. Moreover, red-edge excitation leads to an increased contribution of the shorter lifetime decay component, while time-resolved emission spectra of NBD-PS showed an atypical blue-shift following excitation. This excludes solvent relaxation restrictions as cause for the measured NBD REES and TRES pointing instead to heterogeneous probe transverse location as the origin of these effects. The latter hypothesis was confirmed by molecular dynamics simulations, from which calculated NBD hydration/location heterogeneity correlated with measured fluorescence lifetimes/REES. Altogether, our combination of theory and experiment-based techniques has led to a considerably improved understanding of NBD photophysics and a reinterpretation of its REES in particular. Pracoviště Ústav fyzikální chemie J.Heyrovského Kontakt Michaela Knapová, michaela.knapova@jh-inst.cas.cz, Tel.: 266 053 196 Rok sběru 2017
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