Substrate and temperature dependence of DNA photolyase repair activity examined with ultrafast spectroscopy

Abstract

The kinetics and energetics of the electron transfer reactions occurring in the enzyme substrate complex of DNA photolyase have been studied by transient absorption spectroscopy with picosecond time resolution in the temperature range 275-90 K. The results can be summarized as follows: (i) The lifetime of 1(FADH-)* depends not only on the presence of the substrate but also on its nature, e.g. the electron transfer from 1(FADH-)* to thymine dimers T< >T and to thymine-uracil dimers T< >U is slower by a factor of 2 to 3 compared to that of U< >U and U< >T. This feature is attributed to the enlarged electronic coupling between 1(FADH-)* and the respective dimers in the absence of the methyl group in the C5 position of the 5'-uracil. (ii) The temperature dependence of the quantum yield was studied,and from this the activation energy for the overall repair process has been estimated to be E(A) = 0.45 ± 0.1 eV.