Abstract
His354 and His358, two highly conserved histidines in Xenopus laevis (6-4) photolyase [equivalent to His401 and His405, in Dunaliella salina (6-4) photolyase], are critical for photoreactivation. They act as a base and an acid, respectively. However, the remaining high repair activity when the pH value is higher than the pKa of histidine suggests the involvement of other basic amino acids in photoreactivation. According to the results of in vivo enzyme assay and three-dimension structural model of Dunaliella salina (6-4) photolyase we hypothesized that Lys281 might be involved in the photoreactivation over the pH range from 10.0 to 11.0. To test this, we generated two mutant forms of the (6-4) photolyase, K281G and K281R mutant, by overlap extension polymerase chain reaction, and performed the enzyme assay with these mutants. From these results we conclude that the Lys281, which is highly conserved in (6-4) photolyases, participates in the photoreactivation and acts as an acid to donate a proton to His401 when the environmental pH is higher than the pKa value of histidine.
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Acknowledgments
This study was supported by The National Natural Science Foundation of China (30871321, 30771312, and 30971817), The National Secial Basic Research Projects of China (SB2007FY400-4), and The National Basic Research Program of China (2009CB125910).
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Zhang, F., Xu, H., Cao, Y. et al. Role of Lys281 in the Dunaliella salina (6-4) Photolyase Reaction. Curr Microbiol 62, 146–151 (2011). https://doi.org/10.1007/s00284-010-9687-1
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DOI: https://doi.org/10.1007/s00284-010-9687-1