Synthesis and photophysical properties of fluorophore-labeled abscisic acid

Abstract

The 8'-benzophenone, 8'-dansylhydrazone, 3'-S-(2-ethyldansylamide), and 3'-S-acetamidofluorescein derivatives of the plant hormone abscisic acid (ABA) have been synthesized for use in photoaffinity labeling (the benzophenone derivative) or fluorescence probe experiments and have been spectroscopically characterized. One of the three fluorescent compounds, the 3'-tethered fluorescein derivative, exhibits spectroscopic and photophysical properties which indicate that it could be an excellent fluorescent probe of ABA interactions in vivo. The 3'-tethered fluorescein and ABA moieties do not interact strongly, so that the fluorescence properties of the fluorescein-labelled hormone are very similar to those of fluorescein itself. Measurements of the absorption, emission, and fluorescence excitation spectra, fluorescence quantum yields, and fluorescence decay parameters of this derivative as a function of pH indicate that the photophysics is dominated by ground and excited state prototropic equilibria involving only the fluorescein moiety. The fluorescein dianion is the only significant absorber and emitter at pH > 6.7, whereas only the cation absorbs and emits at pH < 0. In the intervening pH range, strong emission from the monoanion and weak emission from two neutral species, tentatively assigned to the zwitterion and the lactone of the fluorescein moiety, is observed.Key words: abscisic acid, fluorescein, synthesis, photophysics.