Excited-state proton transfer of equilenin and dihydroequilenin: interaction with bilayer vesicles.
The two-state excited-state proton-transfer process for d-equilenin [d-3-hydroxyestra-1,3,-5(10),6,8-pentaen-17-one] and dihydroequilenin is found to depend both on pH and on proton acceptor concentration. Both the protonated and deprotonated forms of the excited molecule are fluorescent. As is the case for 2-naphthol, the excited-state pKa (pKa*) is substantially lower than the ground-state pKa. Fluorescence decay studies have been performed as a function of emission wavelength in aqueous solutions at pH 6.9 in the presence of acetate anion (0.1 M). At this pH, both back-reaction from the excited-state and ground-state heterogeneity are minimal. A monoexponential decay is found in the blue region of the spectrum and a biexponential decay on the red edge. The lifetimes measured across both regions are constant, with a negative preexponential term, characteristic of an excited-state reaction, evident at longer wavelengths. Decay-associated spectra (DAS), the preexponential terms associated with the measured lifetimes, have been acquired for these aqueous solutions. Equilenin and dihydroequilenin are found to adsorb to dimyristoyllecithin (DML) vesicles. Rates for excited-state proton transfer are greatly reduced when dihydroequilenin adsorbs to vesicles. The accessibility of the bound probe to acetate as a proton acceptor depends on the cholesterol content of the vesicles.
D015068:17-Ketosteroids; D002784:Cholesterol; D004134:Dimyristoylphosphatidylcholine; D004855:Equilenin / Q000031:analogs & derivatives; D007700:Kinetics; D008051:Lipid Bilayers; D008433:Mathematics; D013053:Spectrophotometry
D015068:17-Ketosteroids; D008051:Lipid Bilayers; C037874:17-dihydroequilenin; D002784:Cholesterol; D004134:Dimyristoylphosphatidylcholine; D004855:Equilenin