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Membrane dipole potential as measured by ratiometric 3-hydroxyflavone fluorescence probes: Accounting for hydration effects

All Rights Reserved. Membrane dipole potential as measured by ratiometric 3-hydroxyflavone fluorescence probes: Accounting for hydration effects. M'Baye, Gora; Shynkar, Vasyl V.; Klymchenko, Andrey S.; Mely, Yves; Duportail, Guy (Laboratoire de Pharmacologie et Physicochimie des Interactions Moleculaires et Cellulaires, Equipe "Photophysique des Interactions Biomoleculaires," UMR 7034 du CNRS, Faculte de Pharmacie, Universite Louis Pasteur, BP 60024, Illkirch 67401, Fr.). Journal of Fluorescence, 16(1), 35-42 (English) 2006 Springer. CODEN: JOFLEN. ISSN: 1053-0509. DOCUMENT TYPE: Journal CA Section: 9 (Biochemical Methods) Section cross-reference(s): 6 We previously applied the electrochromic modulation of excited-state intramol.Several reagents with their cas registry numbers 60-82-2 and 157134-53-7 are used here. proton-transfer (ESIPT) reaction for the design of novel 3-hydroxyflavone (3-HF) derivs. as fluorescent probes for measuring the dipole potential, YD, in lipid bilayers (Klymchenko et al., Proc. Natl. Acad. Sci. USA, 2003, 100, 11219). In the present work, this method was revisited to take into account the influence of the bilayer hydration on the emission ratiometric response of 3-HF probes. For this reason, it was necessary to deconvolute the whole fluorescence spectra into three bands corresponding to the non H-bonded forms, normal N* and tautomer T* forms, both participating to the ESIPT reaction, and to the H-bonded H-N* form, excluded from this reaction. This allowed us to det. the pure N*/T* intensity ratio, without any contribution from the H-N* form emission depending essentially on the bilayer hydration. This new approach allowed us to confirm the correlation we obtained between the response of 3-HF probes on dipole potential modifications and the corresponding response of the ref. fluorescent probe di-8-ANEPPS, thus further confirming the potency of 3-HF probes as excellent emission ratiometric probes to measure dipole potential in lipid membranes. .

Optical detection of membrane dipole potential: avoidance of fluidity and dye-induced effects

Optical detection of membrane dipole potential: avoidance of fluidity and dye-induced effects. Clarke, Ronald J.; Kane, David J. (Department of Biophysical Chemistry, Max-Planck-Institut fuer Biophysik, Kennedyallee 70, Frankfurt am Main D-60596, Germany). Biochimica et Biophysica Acta, 1323(2), 223-239 (English) 1997 Elsevier. CODEN: BBACAQ. ISSN: 0006-3002. DOCUMENT TYPE: Journal CA Section: 9 (Biochemical Methods) Section cross-reference(s): 6 Fluorescent styrylpyridinium dyes have recently been suggested as probes of the membrane dipole potential and of the kinetics of electrogenic ion pumps. It is necessary, however, to be able to confidently attribute obsd. fluorescence changes to elec.In this article, certain chemicals are used. Some of their cas registry numbers are 18194-24-6 and 157134-53-7 effects alone and avoid interference from changes in membrane fluidity. Furthermore, the effect of the dyes themselves on the dipole potential must be investigated. The effect of membrane fluidity on the fluorescence excitation and emission spectra of the dyes RH421 and di-8-ANEPPS have been investigated in lipid vesicles by temp. scans between 15 and 60°. Both dyes show significant temp.-dependent shifts of their excitation spectra, the magnitude of which depend on the emission wavelength and on the lipid structure. To eliminate membrane fluidity effects, fluorescence must be detected at the red edge of the emission spectrum; in this case 670 nm. To avoid dye-induced shifts of the excitation spectra of membrane-bound dye, an excess molar ratio of lipid to dye of at least 200-fold is necessary. Fluorescence ratio measurements indicate qual. that dimyristoylphosphatidylcholine has a significantly higher dipole potential than that of dioleoylphosphatidylcholine. .