title: Kinetic and thermodynamic analysis of the light-induced processes in plant and cyanobacterial phytochromes.
abstract: The light-induced processes of the biological photoreceptor phytochrome (recombinant phyA of oat and recombinant CphA from the cyanobacterium Tolypothrix PCC7601) have been investigated in a time-resolved manner in the temperature range from 0 to 30°C. Both proteins were heterologously expressed and assembled in vitro with phycocyanobilin. The Pr state of plant phytochrome phyA is converted to the Pfr state after formation of four intermediates with an overall quantum yield of ~18%. The reversal reaction (Pfr-to-Pr) shows several intermediates, all of which, even the first detectable one, exhibit already all spectral features of the Pr state. The canonical phytochrome CphA from Tolypothrix showed a similar intermediate sequence as its plant ortholog. Whereas the kinetics for the forward reaction (Pr-to-Pfr) was nearly identical for both proteins, the reverse process (Pr formation) in the cyanobacterial phytochrome was slower by a factor of three. As found for the Pfr-to-Pr intermediates in the plant protein, also in CphA all detectable intermediates showed the spectral features of the Pr form. For both phytochromes, activation parameters for both the forward and the backward reaction pathways were determined.
authors.affiliation: Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Igor
authors.initials: I
authors.lastname: Chizhov
authors.forename: Björn
authors.initials: B
authors.lastname: Zorn
authors.forename: Dietmar J
authors.initials: DJ
authors.lastname: Manstein
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Gärtner
mesh_terms: D018554:Avena; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D000458:Cyanobacteria; D007700:Kinetics; D008027:Light; D055668:Photochemical Processes / Q000528:radiation effects; D010834:Phytochrome / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*; D013696:Temperature; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D010834:Phytochrome