Biochemical and spectroscopic characterization of the bacterial phytochrome of Pseudomonas aeruginosa.
Phytochromes are photochromic biliproteins found in plants as well as in some cyanotrophic, photoautotrophic and heterotrophic bacteria. In many bacteria, their function is largely unknown. Here we describe the biochemical and spectroscopic characterization of recombinant bacterial phytochrome from the opportunistic pathogen Pseudomonas aeruginosa (PaBphP). The recombinant protein displays all the characteristic features of a bonafide phytochrome. In contrast with cyanobacteria and plants, the chromophore of this bacterial phytochrome is biliverdin IXalpha, which is produced by the heme oxygenase BphO in P. aeruginosa. This chromophore was shown to be covalently attached via its A-ring endo-vinyl group to a cysteine residue outside the defined bilin lyase domain of plant and cyanobacterial phytochromes. Site-directed mutagenesis identified Cys12 and His247 as being important for chromophore binding and photoreversibility, respectively. PaBphP is synthesized in the dark in the red-light-absorbing Pr form and immediately converted into a far-red-light-absorbing Pfr-enriched form. It shows the characteristic red/far-red-light-induced photoreversibility of phytochromes. A chromophore analog that lacks the C15/16 double bond was used to show that this photoreversibility is due to a 15Z/15E isomerization of the biliverdin chromophore. Autophosphorylation of PaBphP was demonstrated, confirming its role as a sensor kinase of a bacterial two-component signaling system.
Tasler
Ronja
R
Institute for Microbiology, Technical University Braunschweig, Germany.
Moises
Tina
T
Frankenberg-Dinkel
Nicole
N
D001664:Biliverdine / Q000737:chemistry / Q000378:metabolism; D007536:Isomerism; D010766:Phosphorylation; D010777:Photochemistry; D010834:Phytochrome / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000737:chemistry* / Q000235:genetics; D013050:Spectrometry, Fluorescence
D010834:Phytochrome; D001664:Biliverdine