Isolation and characterization of a new peroxiredoxin from poplar sieve tubes that uses either glutaredoxin or thioredoxin as a proton donor.
A sequence coding for a peroxiredoxin (Prx) was isolated from a xylem/phloem cDNA library from Populus trichocarpa and subsequently inserted into an expression plasmid yielding the construction pET-Prx. The recombinant protein was produced in Escherichia coli cells and purified to homogeneity with a high yield. The poplar Prx is composed of 162 residues, a property that makes it the shortest plant Prx sequence isolated so far. It was shown that the protein is monomeric and possesses two conserved cysteines (Cys). The Prx degrades hydrogen peroxide and alkyl hydroperoxides in the presence of an exogenous proton donor that can be either thioredoxin or glutaredoxin (Grx). Based on this finding, we propose that the poplar protein represents a new type of Prx that differs from the so-called 2-Cys and 1-Cys Prx, a suggestion supported by the existence of natural fusion sequences constituted of a Prx motif coupled to a Grx motif. The protein was shown to be highly expressed in sieve tubes where thioredoxin h and Grx are also major proteins.
Rouhier
N
N
Unité Mixte de Recherche Interaction Arbres Microorganisms, Institut National de la Recherche Agronomique-Université Henri Poincaré Nancy I. Biochimie et Biologie Moléculaire Végétales, Université Henri Poincaré, 54506 Vandoeuvre cedex, France.
Gelhaye
E
E
Sautiere
P E
PE
Brun
A
A
Laurent
P
P
Tagu
D
D
Gerard
J
J
de Faÿ
E
E
Meyer
Y
Y
Jacquot
J P
JP
D000595:Amino Acid Sequence; D001693:Biological Transport, Active; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D054477:Glutaredoxins; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D009195:Peroxidase / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D054464:Peroxiredoxins; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism*; D011522:Protons; D031308:Salicaceae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D016415:Sequence Alignment; D013438:Sulfhydryl Compounds / Q000032:analysis; D013879:Thioredoxins / Q000378:metabolism*
D054477:Glutaredoxins; D011506:Proteins; D011522:Protons; D013438:Sulfhydryl Compounds; D013879:Thioredoxins; D010088:Oxidoreductases; D010544:Peroxidases; D054464:Peroxiredoxins; D009195:Peroxidase