Publication 641

J. Am. Chem. Soc. 131, 426-427, 2009. DOI: 10.1021/ja806540j
What Makes the Difference between a Cryptochrome and DNA Photolyase ? A Spectroelectrochemical Comparison of the Flavin Redox Transitions.

Laboratoire d‘Electrochimie Moléculaire, UMR CNRS 7591 Université Paris Diderot, 2 place Jussieu 75251 Paris Cedex 05 (France), CEA, IBITECS, Laboratoire de Photocatalyse et Biohydrogène, Gif sur Yvette, F-91191, France, CNRS, URA2096, Gif sur Yvette, F-91191, France, Department of Cell Biology and Genetics, MGC, Erasmus University Medical Centre, PO Box 2040, Rotterdam, 3000 CA, The Netherlands;, Univ Paris 06, 4 Pl Jussieu Casier 156, F-75005 Paris, France, and Penn State University, Philadelphia, Pennsylvania 19104

Cryptochromes and DNA photolyases are highly homologous flavoproteins that accomplish completely different tasks. While plant cryptochrome1 functions as blue light photoreceptor that triggers various morphogenic reactions, photolyases repair UV-induced DNA damages. Both enzymes share the photoactive cofactor, noncovalently bound FAD. For photolyase, the reaction mechanism involves electron transfer to the substrate from the excited-state of fully reduced flavin. For cryptochrome, photoexcitation of the oxidized flavin leads to formation of the semireduced radical FADH•. Key parameters for the redox state of the flavin in the cell are the midpoint potentials E1 and E2 for the oxidized/semireduced and semireduced/fully reduced transitions, respectively. A link between cryptochrome function and its cofactor's redox states has been suggested early on, but no reliable determinations of midpoint potentials have been available. Here we report spectroelectrochemical titrations of cryptochrome1 from Arabidopsis thaliana and photolyases from both E. coli and Anacystis nidulans at pH 7.4. For the cryptochrome, we obtained E1 ˜ E2 ˜ -160 mV vs NHE, strongly deviating from the photolyases where FADH• could not be oxidized up to 400 mV, and E2 ˜ -40 mV. Functional and evolutionary implications are discussed, highlighting the role of an asparagine-to-aspartate replacement close to N5 of the flavin.