Interaction of Glutathione S-Transferase with Hypericin: A Photophysical Study

The photophysics of hypericin have been studied in its complex with two different isoforms, A1-1 and P1-1, of the protein glutathione S-transferase (GST). One molecule of hypericin binds to each of the two GST subunits. Comparisons are made with our previous results for the hypericin/human serum albumin complex (Photochem. Photobiol. 1999, 69, 633−645). Hypericin binds with high affinity to the GSTs:  0.65 μM for the A1-1 isoform and 0.51 μM for the P1-1 isoform (Biochemistry 2004, 43, 12761−12769). The photophysics and activity of hypericin are strongly modulated by the binding protein. Intramolecular hydrogen-atom transfer is suppressed in both cases. Most importantly, while there is significant singlet oxygen generation from hypericin bound to GST A1-1, binding to GST P1-1 suppresses singlet oxygen generation to almost negligible levels. The data are rationalized in terms of a simple model in which the hypericin photophysics depends entirely upon the decay of the triplet state by two competing processes, quenching by oxygen to yield singlet oxygen and ionization, the latter of these two are proposed to be modulated by A1-1 and P1-1

Role of Solvent in Excited-State Proton Transfer in Hypericin

The excited-state proton transfer of hypericin is monitored by the rise time (-6-1 2 ps in the solvents investigated) of the component of stimulated emission corresponding to the formation of the long-lived (-5 ns) fluorescent tautomer. The assignment of this excited-state process to proton transfer has been verified by noting that a hypericin analog (mesonaphthobianthrone) lacking labile protons is not fluorescent unless its carbonyl groups are protonated. Recent experimental studies on other systems have suggested that three solvent properties play important roles in excited-state proton transfer: viscosity, hydrogen-bonding character, and dynamic solvation. We find that for hypericin, in a range of protic, aprotic, hydrogen-bonding, and non-hydrogen-bonding solvents in which the viscosity changes by a factor of 60 and the average solvation time changes by a factor of 100, the excited-state proton-transfer rate of hypericin is uncorrelated with these properties and varies not more than a factor of 2 (- 6-1 2 ps) at room temperature. The relative contribution of the bulk solvent polarity is considered, and the role of intramolecular vibrations of hypericin on the proton-transfer rate is discussed

Monophotonic Ionization of 7-Azaindole, Indole, and Their Derivatives and the Role of Overlapping Excited States

7-Azaindole undergoes monophotonic ionization just as its counterpart, indole. This result suggests that 7-azaindole is qualitatively more similar to indole than has previously been recognized. The appearance of the solvated electron for zwitterionic and anionic 7-azatryptophan and for 7-azaindole in water and methanol is complete within 1 ps, which indicates that the fluorescent state whose lifetime is \u3elo0 ps cannot be the source of the electron. The origin of the electron is related to the presence of closely spaced or overlapping excited states in 7-azaindole, which is another similarity that this chromophore bears with respect to indole. The fluorescence quantum yield of 7-azaindole is shown to be excitation wavelength dependent. The excitation-wavelength dependence and the temperaturedependence of the fluorescence quantum yield of 7-azaindole are explored and related to the production of the solvated electron. The implications of these observations for the use of 7-azatryptophan as an alternative to tryptophan as a probe of protein structure and dynamics are discussed