Assembly of plant ferredoxin-NADP+ oxidoreductase in Escherichia coli requires GroE molecular chaperones.
We have recently reported the expression in Escherichia coli of an enzymatically competent ferredoxin-NADP+ oxidoreductase from cloned pea genes encoding either the mature enzyme or its precursor protein (Ceccarelli, E. A., Viale, A. M., Krapp, A. R., and Carrillo, N. (1991) J. Biol. Chem. 266, 14283-14287). Processing to the mature form by bacterial protease(s) and FAD assembly occurred in the bacterial cytosol. Expression of ferredoxin-NADP+ reductase in chaperonin-deficient (groE-) mutants of E. coli resulted in partial reductase assembly at permissive growth temperatures (i.e. 30 degrees C), and in total breakdown of holoenzyme assembly, and accumulation as aggregated inclusion bodies at non-permissive temperatures (i.e. 42 degrees C). Coexpression in these mutants of a cloned groESL operon from the phototrophic bacterium Chromatium vinosum resulted in partial or total recoveries of ferredoxin-NADP+ reductase assembly. The overall results indicate that bacterial chaperonins are required for the productive folding/assembly of eucaryotic ferredoxin-NADP+ reductase expressed in E. coli.
Departamento de Ciencias Biológicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina.
N
N
Carrillo
E A
EA
Ceccarelli
A R
AR
Krapp
S
S
Boggio
R G
RG
Ferreyra
A M
AM
Viale
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018835:Chaperonin 10; D018834:Chaperonin 60; D018833:Chaperonins / Q000378:metabolism*; D002844:Chromatium / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005287:Ferredoxin-NADP Reductase / Q000235:genetics*; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000235:genetics; D009876:Operon; D010957:Plasmids
D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; C028715:GroE protein, E coli; C097970:GroESL protein, Bacteria; D006360:Heat-Shock Proteins; D005287:Ferredoxin-NADP Reductase; D018833:Chaperonins