Parts | Days | Selection | Search | Downloads | Help
BP: Fachverband Biologische Physik
BP 29: Biomolecular Spectroscopy
BP 29.9: Talk
Thursday, March 25, 2010, 16:45–17:00, H45
Photocycle Dynamics of the E149A Mutant of Cryptochrome 3 from Arabidopsis thaliana — Peyman Zirak1, •Alfons Penzkofer1, Judit Moldt2, Richard Pokorny2, Alfred Batschauer2, and Lars-Oliver Essen3 — 1Institut II - Experimentelle und Angewandte Physik, Universität Regensburg, Universitätsstr. 31, 93053 Regensburg — 2Fachbereich Biologie, Pflanzenphysiologie/Photobiologie, Philipps-Universität, Karl-von-Frisch-Str. 8, 35032 Marburg — 3Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Str., 35032 Marburg
The E149A mutant of the cryDASH member cryptochrome 3 (cry3) from Arabidopsis thaliana was characterized in vitro by absorption and emission spectroscopy. The mutant protein non-covalently binds the cofactor flavin adenine dinucleotide (FAD), but not the second cofactor 5,10-methenyl-tetrahydrofolate (MTHF). Thus, the photo-dynamics caused by FAD is accessible without the intervening coupling with MTHF. In dark adapted cry3-E149A, FAD is present in the oxidized form (FADox), semiquinone form (FADH.), and anionic hydroquinone form (FADredH−). Blue-light photo-excitation of previously unexposed cry3-E149A transfers FADox to the anionic semiquinone form (FAD.−) with a quantum efficiency of 0.02 and a back recovery time of 10 s (photocycle I). Prolonged photo-excitation leads to an irreversible protein re-conformation leading to a change in the photo-cycle dynamics with photo-conversion of FADox to FADH. (efficiency 0.00032), of FADH. to FADredH− (efficiency 0.016), and thermal back equilibration in the dark on a minute timescale (photocycle II).