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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Autor principal:	Moreno de Colonna, Silvia
Publicado:	2012
Materias:	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 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
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00320781_v53_n6_p1144_Alvarez http://hdl.handle.net/20.500.12110/paper_00320781_v53_n6_p1144_Alvarez
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_00320781_v53_n6_p1144_Alvarez2023-06-08T14:59:54Z Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity Moreno de Colonna, Silvia 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. 2012 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00320781_v53_n6_p1144_Alvarez 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 Moreno de Colonna, Silvia 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.
author	Moreno de Colonna, Silvia
author_facet	Moreno de Colonna, Silvia
author_sort	Moreno de Colonna, Silvia
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
publishDate	2012
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00320781_v53_n6_p1144_Alvarez http://hdl.handle.net/20.500.12110/paper_00320781_v53_n6_p1144_Alvarez
work_keys_str_mv	AT morenodecolonnasilvia functionalcharacterizationofresiduesinvolvedinredoxmodulationofmaizephotosyntheticnadpmalicenzymeactivity
_version_	1768544585138569216

Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-Malic enzyme activity

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