Investigation of molecular mechanisms in photodynamic action and radiobiology with nanosecond flash photolysis and pulse radiolysis. Progress report, July 1, 1975--June 30, 1976

Flash photolysis studies have shown that exposed tyrosyl or tryptophyl residues of enzymes are photoionized at approximately the same quantum efficiency as the aqueous amino acids. Comparisons with permanent damage indicate that this primary process contributes to inactivation when a photolyzed aromatic residue is essential or is located adjacent to a catalytic, non-chromoporic residue. Studies on sensitization by the phototherapeutic drug, 8-methoxypsoralen (8-MOP), show the triplet state can lead to singlet oxygen generation and that weak binding to poly (dA-dT) suppresses this process. 8-MOP was found to be potent sensitizer of yeast photoinactivation with negligible post-irradiation recovery. Spinlable ESR measurements made on yeast cells after photodynamic treatment show that the membrane damage generated by the penetrating dye Toluidine Blue 0 is more extensive than the external dye, Eosin Y. In related ionizing radiation studies, it was found that 8-MOP is an anoxic radiosensitizer of T7 phage, attributed to enhancement of crosslinks; that complexing of eosin to lysozyme sensitizes the enzyme to oxidizing radicals; that the reactions of Br2- and (CNS)2- with tryptophan generate the same initial radical as uv-excited photoionization