Zirak, Peyman and Penzkofer, Alfons and Schiereis, T. and Hegemann, P. and Jung, A. and Schlichting, I. (2006) Photodynamics of the small BLUF protein BlrB from Rhodobacter sphaeroides. Journal of Photochemistry and Photobiology B: Biology 83 (3), pp. 180-194.
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The BLUF protein BlrB from the non-sulphur anoxyphototrophic purple bacterium Rhodobacter sphaeroides is characterized by absorption and emission spectroscopy. BlrB expressed from E. coli binding FAD, FMN, and riboflavin (called BrlBI) and recombinant BlrB containing only FAD (called BlrBII) are investigated. The dark-adapted proteins exist in two different receptor conformations (receptor states) with different sub-nanosecond fluorescence lifetimes (BLUFr,f and BLUFr,sl). Some of the flavin-cofactor (ca. 8%) is unbound in thermodynamic equilibrium with the bound cofactor. The two receptor conformations are transformed to putative signalling states (BLUFs,f and BLUFs,sl) of decreased fluorescence efficiency and shortened fluorescence lifetime by blue-light excitation. In the dark at room temperature both signalling states recover back to the initial receptor states with a time constant of about 2 s. Quantum yields of signalling state formation of about 90% for BlrBII and about 40% for BlrBI were determined by intensity dependent transmission measurements. Extended blue-light excitation causes unbound flavin degradation (formation of lumichrome and lumiflavin-derivatives) and bound cofactor conversion to the semiquinone form. The flavin-semiquinone further reduces and the reduced flavin re-oxidizes back in the dark. A photo-dynamics scheme is presented and relevant quantum efficiencies and time constants are determined.
|Institutions:||Physics > Institute of Experimental and Applied Physics > Retired Professors > Group Alfons Penzkofer|
|Keywords:||BLUF domain; BlrB from Rhodobacter sphaeroides; Blue-light photoreceptor; Absorption spectroscopy; Fluorescence spectroscopy; Photo-cycle; Photo-degradation; Photo-reduction; Flavoprotein; Flavins; FAD; Flavin-semiquinone|
|Subjects:||500 Science > 530 Physics|
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Yes|
|Deposited On:||20 Mar 2007|
|Last Modified:||20 Jul 2011 21:01|