Engineering functional artificial hybrid proteins between poplar peroxiredoxin II and glutaredoxin or thioredoxin.
The existence of natural peroxiredoxin-glutaredoxin hybrid enzymes in several bacteria is in line with previous findings indicating that poplar peroxiredoxin II can use glutaredoxin as an electron donor. This peroxiredoxin remains however unique since it also uses thioredoxin with a quite good efficiency. Based on the existing fusions, we have created artificial enzymes containing a poplar peroxiredoxin module linked to glutaredoxin or thioredoxin modules. The recombinant fusion enzymes folded properly into non-covalently bound homodimers or homotetramers. Two of the three protein constructs exhibit peroxidase activity, a reaction where the two modules need to function together, but they also display enzymatic activities specific of each module. In addition, mass spectrometry analyses indicate that the Prx module can be both glutathiolated or overoxidized in vitro. This is discussed in the light of the Prx reactivity.
UMR 1136 Interactions Arbres Mircoorganismes INRA-UHP, IFR 110 GEEF, Faculté des Sciences, 54506 Vandouevre-les-Nancy Cedex, France. nrouhier@scbiol.uhp-nancy.fr
Nicholas
N
Rouhier
Filipe
F
Gama
Gunnar
G
Wingsle
Eric
E
Gelhaye
Pierre
P
Gans
Jean-Pierre
JP
Jacquot
D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D054477:Glutaredoxins; D005978:Glutathione / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism; D054464:Peroxiredoxins; D032107:Populus / Q000201:enzymology*; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics* / Q000378:metabolism
D054477:Glutaredoxins; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins; D010088:Oxidoreductases; D010544:Peroxidases; D054464:Peroxiredoxins; D005978:Glutathione