Recombinant holophytochrome in Escherichia coli.
We have successfully co-expressed two genes from the bilin biosynthetic pathway of Synechocystis together with cyanobacterial phytochrome 1 (Cph1) from the same organism to produce holophytochrome in Escherichia coli. Heme oxygenase was used to convert host heme to biliverdin IXalpha which was then reduced to phycocyanobilin via phycocyanobilin:ferredoxin oxidoreductase, presumably with the aid of host ferredoxin. In this host environment Cph1 apophytochrome was able to autoassemble with the phycocyanobilin in vivo to form fully photoreversible holophytochrome. The system can be used as a tool for further genetic studies of phytochrome function and signal transduction as well as providing an excellent source of holophytochrome for physicochemical studies.
Landgraf
F T
FT
Plant Physiology, Justus-Leibig-University Giessen, Germany.
Forreiter
C
C
Hurtado Picó
A
A
Lamparter
T
T
Hughes
J
J
D000818:Animals; D001426:Bacterial Proteins; D001654:Bile Pigments / Q000096:biosynthesis; D000458:Cyanobacteria / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D020283:Photoreceptors, Microbial; D052980:Phycobilins; D010798:Phycocyanin / Q000096:biosynthesis / Q000235:genetics; D010834:Phytochrome / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011494:Protein Kinases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011758:Pyrroles; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D013057:Spectrum Analysis; D013696:Temperature; D045725:Tetrapyrroles; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D001654:Bile Pigments; D029968:Escherichia coli Proteins; D020283:Photoreceptors, Microbial; D052980:Phycobilins; D011758:Pyrroles; D011994:Recombinant Proteins; D045725:Tetrapyrroles; D010798:Phycocyanin; D010834:Phytochrome; C011069:phycocyanobilin; D010088:Oxidoreductases; C442236:phycocyanobilin ferredoxin oxidoreductase, Synechocystis; D006419:Heme Oxygenase (Decyclizing); D011494:Protein Kinases; C413623:Cph1 phytochrome protein, bacteria