Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity
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, the...
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00320781_v53_n6_p1144_Alvarez |
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todo:paper_00320781_v53_n6_p1144_Alvarez2023-10-03T14:44:49Z Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity Alvarez, C.E. Detarsio, E. Moreno, S. Andreo, C.S. Drincovich, M.F. 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 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. Fil:Moreno, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00320781_v53_n6_p1144_Alvarez |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 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 |
| topic_facet |
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 |
| description |
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. |
| format |
JOUR |
| author |
Alvarez, C.E. Detarsio, E. Moreno, S. Andreo, C.S. Drincovich, M.F. |
| author_facet |
Alvarez, C.E. Detarsio, E. Moreno, S. Andreo, C.S. Drincovich, M.F. |
| author_sort |
Alvarez, C.E. |
| title |
Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity |
| title_short |
Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity |
| title_full |
Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity |
| title_fullStr |
Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity |
| title_full_unstemmed |
Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity |
| title_sort |
functional characterization of residues involved in redox modulation of maize photosynthetic nadp-malic enzyme activity |
| url |
http://hdl.handle.net/20.500.12110/paper_00320781_v53_n6_p1144_Alvarez |
| work_keys_str_mv |
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1807320183505682432 |