Functional and structural aspects of poplar cytosolic and plastidial type A methionine sulfoxide reductases
2007
Rouhier , Nicolas (INRA (France). UMR 1136 Interactions Arbres/Micro-organismes) | Kauffmann , Brice (Centre National de la Recherche Scientifique, Vandoeuvre cedex(France). UMR 7036 CNRS-UHP, Nancy Universite´s, LCM3B, Groupe de Biocristallographie, Faculte´ des Sciences et Techniques, BP 239) | Tete-Favier , Frédérique (Centre National de la Recherche Scientifique, Vandoeuvre cedex(France). UMR 7036 CNRS-UHP, Nancy Universités, LCM3B, Groupe de Biocristallographie,Faculte´ des Sciences et Techniques) | Palladino , Pasquale (Centre National de la Recherche Scientifique, Vandoeuvre cedex(France). UMR 7036 CNRS-UHP, Nancy Universités, LCM3B, Groupe de Biocristallographie, Faculte´ des Sciences et Techniques, BP 239) | Gans , Pierre (Commissariat à l'Energie Atomique et aux Energies Alternatives, Grenoble Cedex 1(France). Laboratoire de Résonance Magnétique Nucléaire, CEA-CNRS-UJF “Jean-Pierre Ebel) | Branlant , Guy (Centre National de la Recherche Scientifique, Vandoeuvre cedex(France). UMR7567 CNRS-UHP, Nancy Universités, Laboratoire MAEM, Groupe Enzymologie Mole´culaire, Faculté des Sciences et Techniques, BP 239) | Jacquot , Jean-Pierre (INRA (France). UMR 1136 Interactions Arbres/Micro-organismes) | Boschi-Muller , Sandrine (Centre National de la Recherche Scientifique, Vandoeuvre cedex(France). UMR7567 CNRS-UHP, Nancy Universités, Laboratoire MAEM, Groupe Enzymologie Moléculaire, Faculté des Sciences et Techniques, BP 239)
The genome of Populus trichocarpa contains five methionine sulfoxide reductase A genes. Here, both cytosolic (cMsrA) and plastidial (pMsrA) poplar MsrAs were analyzed. The two recombinant enzymes are active in the reduction of methionine sulfoxide with either dithiothreitol or poplar thioredoxin as a reductant. In both enzymes, five cysteines, at positions 46, 81, 100, 196, and 202, are conserved. Biochemical and enzymatic analyses of the cysteine-mutated MsrAs support a catalytic mechanism involving three cysteines at positions 46, 196, and 202. Cys46 is the catalytic cysteine, and the two C-terminal cysteines, Cys196 and Cys202, are implicated in the thioredoxin-dependent recycling mechanism. Inspection of the pMsrA x-ray three-dimensional structure, which has been determined in this study, strongly suggests that contrary to bacterial and Bos Taurus MsrAs, which also contain three essential Cys, the last C-terminal Cys202, but not Cys196, is the first recycling cysteine that forms a disulfide bond with the catalytic Cys46. Then Cys202 forms a disulfide bond with the second recycling cysteine Cys196 that is preferentially reduced by thioredoxin. In agreement with this assumption, Cys202 is located closer to Cys46 compared with Cys196 and is included in a 202CYG204 signature specific for most plant MsrAs. The tyrosine residue corresponds to the one described to be involved in substrate binding in bacterial and B. taurus MsrAs. In these MsrAs, the tyrosine residue belongs to a similar signature as found in plant MsrAs but with the first C-terminal cysteine instead of the last C-terminal cysteine.
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