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Two highly similar plastidic NADP-malic enzymes (NADP-MEs) are found in the C 4 species maize (Zea mays); one exclusively expressed in the bundle sheath cells (BSCs) and involved in C 4 photosynthesis (ZmC 4-NADP-ME); and the other (ZmnonC 4-NADP-ME) with housekeeping roles. In the present work, these two NADP-MEs were analyzed regarding their redox-dependent activity modulation. The results clearly show that ZmC 4-NADP-ME is the only one modulated by redox status, and that its oxidation produces a conformational change limiting the catalytic process, although inducing higher affinity binding of the substrates. The reversal of ZmC 4-NADP-ME oxidation by chemical reductants suggests the presence of thiol groups able to form disulfide bonds. In order to identify the cysteine residues involved in the activity modulation, site-directed mutagenesis and MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) analysis of ZmC 4-NADP-ME were performed. The results obtained allowed the identification of Cys192, Cys246 (not conserved in ZmnonC 4-NADP-ME), Cys270 and Cys410 as directly or indirectly implicated in ZmC 4-NADP-ME redox modulation. These residues may be involved in forming disulfide bridge(s) or in the modulation of the oxidation of critical residues. Overall, the results indicate that, besides having acquired a high level of expression and localization in BSCs, ZmC 4-NADP-ME displays a particular redox modulation, which may be required to accomplish the C 4 photosynthetic metabolism. Therefore, the present work could provide new insights into the regulatory mechanisms potentially involved in the recruitment of genes for the C 4 pathway during evolution. © 2012 The Author.


Documento: Artículo
Título:Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity
Autor:Alvarez, C.E.; Detarsio, E.; Moreno, S.; Andreo, C.S.; Drincovich, M.F.
Filiación:Centro de Estudios Fotosintéticos y Bioquímicos (CEFOBI), Universidad Nacional de Rosario, Suipacha 531, Rosario, Argentina
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, 1428 Buenos Aires, Argentina
Palabras clave:C 4 photosynthesis; Maize; NADP-malic enzyme; Redox modulation; Structure-function relationship; chloroplast protein; cysteine; malate dehydrogenase; malate dehydrogenase (oxaloacetate decarboxylating) (NADP+); malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+); recombinant protein; vegetable protein; amino acid sequence; article; chloroplast; enzyme activation; enzyme specificity; enzymology; Escherichia coli; genetics; maize; mass spectrometry; metabolism; methodology; molecular genetics; mutation; oxidation reduction reaction; photosynthesis; sequence alignment; site directed mutagenesis; structure activity relation; Amino Acid Sequence; Chloroplast Proteins; Chloroplasts; Cysteine; Enzyme Activation; Escherichia coli; Malate Dehydrogenase; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Oxidation-Reduction; Photosynthesis; Plant Proteins; Recombinant Proteins; Sequence Alignment; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure-Activity Relationship; Substrate Specificity; Zea mays
Página de inicio:1144
Página de fin:1153
Título revista:Plant and Cell Physiology
Título revista abreviado:Plant Cell Physiol.
CAS:cysteine, 4371-52-2, 52-89-1, 52-90-4; malate dehydrogenase, 9001-64-3; Chloroplast Proteins; Cysteine, 52-90-4; Malate Dehydrogenase,; Plant Proteins; Recombinant Proteins; malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+),


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---------- APA ----------
Alvarez, C.E., Detarsio, E., Moreno, S., Andreo, C.S. & Drincovich, M.F. (2012) . Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity. Plant and Cell Physiology, 53(6), 1144-1153.
---------- CHICAGO ----------
Alvarez, C.E., Detarsio, E., Moreno, S., Andreo, C.S., Drincovich, M.F. "Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity" . Plant and Cell Physiology 53, no. 6 (2012) : 1144-1153.
---------- MLA ----------
Alvarez, C.E., Detarsio, E., Moreno, S., Andreo, C.S., Drincovich, M.F. "Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity" . Plant and Cell Physiology, vol. 53, no. 6, 2012, pp. 1144-1153.
---------- VANCOUVER ----------
Alvarez, C.E., Detarsio, E., Moreno, S., Andreo, C.S., Drincovich, M.F. Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity. Plant Cell Physiol. 2012;53(6):1144-1153.