Identification of plant glutaredoxin targets.
Glutaredoxins (Grxs) are small ubiquitous proteins of the thioredoxin (Trx) family, which catalyze dithiol-disulfide exchange reactions or reduce protein-mixed glutathione disulfides. In plants, several Trx-interacting proteins have been isolated from different compartments, whereas very few Grx-interacting proteins are known. We describe here the determination of Grx target proteins using a mutated poplar Grx, various tissular and subcellular plant extracts, and liquid chromatography coupled to tandem mass spectrometry detection. We have identified 94 putative targets, involved in many processes, including oxidative stress response [peroxiredoxins (Prxs), ascorbate peroxidase, catalase], nitrogen, sulfur, and carbon metabolisms (methionine synthase, alanine aminotransferase, phosphoglycerate kinase), translation (elongation factors E and Tu), or protein folding (heat shock protein 70). Some of these proteins were previously found to interact with Trx or to be glutathiolated in other organisms, but others could be more specific partners of Grx. To substantiate further these data, Grx was shown to support catalysis of the stroma beta-type carbonic anhydrase and Prx IIF of Arabidopsis thaliana, but not of poplar 2-Cys Prx. Overall, these data suggest that the interaction could occur randomly either with exposed cysteinyl disulfide bonds formed within or between target proteins or with mixed disulfides between a protein thiol and glutathione.
Rouhier
Nicolas
N
Unité Mixte de Recherches 1136 INRA UHP (Interaction Arbres Microorganismes), IFR 110 Génomique Ecophysiologie et Ecologie Fonctionnelles, Université Henri Poincaré, Vandoeuvre, France. nrouhier@scbiol.uhp-nancy.fr
Villarejo
Arsenio
A
Srivastava
Manoj
M
Gelhaye
Eric
E
Keech
Olivier
O
Droux
Michel
M
Finkemeier
Iris
I
Samuelsson
Göran
G
Dietz
Karl Josef
KJ
Jacquot
Jean-Pierre
JP
Wingsle
Gunnar
G
D000255:Adenosine Triphosphate / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology / Q000378:metabolism*; D002245:Carbon Dioxide / Q000378:metabolism; D019069:Cell Respiration / Q000528:radiation effects; D019343:Citric Acid / Q000378:metabolism; D004579:Electron Transport; D054477:Glutaredoxins; D018832:Molecular Chaperones / Q000378:metabolism; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism; D054464:Peroxiredoxins; D010777:Photochemistry; D011134:Polysaccharides / Q000096:biosynthesis / Q000378:metabolism; D014176:Protein Biosynthesis; D013455:Sulfur / Q000378:metabolism
D054477:Glutaredoxins; D018832:Molecular Chaperones; D011134:Polysaccharides; D002245:Carbon Dioxide; D019343:Citric Acid; D013455:Sulfur; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D010544:Peroxidases; D054464:Peroxiredoxins; D010100:Oxygen