PoplarV1, Main, Exploration, bibRecord, 002701

Enzymatic properties of Populus α- and β-NAD-ME recombinant proteins.

Identifieur interne : 002701 ( Main/Exploration ); précédent : 002700; suivant : 002702

Enzymatic properties of Populus α- and β-NAD-ME recombinant proteins.

Auteurs : Jinwen Liu [République populaire de Chine] ; Qiguo Yu ; Nabil I. Elsheery ; Yuxiang Cheng

Source :

International journal of molecular sciences [ 1422-0067 ] ; 2013.

RBID : pubmed:23797660

Descripteurs français

KwdFr :
- Cinétique (MeSH), Escherichia coli (métabolisme), NAD (métabolisme), Populus (métabolisme), Protéines recombinantes (génétique).
MESH :
- génétique : Protéines recombinantes.
- métabolisme : Escherichia coli, NAD, Populus.
- Cinétique.

English descriptors

KwdEn :
- Escherichia coli (metabolism), Kinetics (MeSH), NAD (metabolism), Populus (metabolism), Recombinant Proteins (genetics).
MESH :
- chemical , genetics : Recombinant Proteins.
- chemical , metabolism : NAD.
- metabolism : Escherichia coli, Populus.
- Kinetics.

Abstract

Plant mitochondrial NAD-malic enzyme (NAD-ME), which is composed of α- and β-subunits in many species, participates in many plant biosynthetic pathways and in plant respiratory metabolism. However, little is known about the properties of woody plant NAD-MEs. In this study, we analyzed four NAD-ME genes (PtNAD-ME1 through PtNAD-ME4) in the genome of Populus trichocarpa. PtNAD-ME1 and -2 encode putative α-subunits, while PtNAD-ME3 and -4 encode putative β-subunits. The Populus NAD-MEs were expressed in Escherichia coli cells as GST-tagged fusion proteins. Each recombinant GST-PtNAD-ME protein was purified to near homogeneity by glutathione-Sepharose 4B affinity chromatography. Milligram quantities of each native protein were obtained from 1 L bacterial cultures after cleavage of the GST tag. Analysis of the enzymatic properties of these proteins in vitro indicated that α-NAD-MEs are more active than β-NAD-MEs and that α- and β-NAD-MEs presented different kinetic properties (Vmax, kcat and kcat/Km). The effect of different amounts of metabolites on the activities of Populus α- and β-NAD-MEs was assessed in vitro. While none of the metabolites evaluated in our assays activated Populus NAD-ME, oxalacetate and citrate inhibited all α- and β-NAD-MEs and glucose-6-P and fructose inhibited only the α-NAD-MEs.

DOI: 10.3390/ijms140712994
PubMed: 23797660
PubMed Central: PMC3742170

Affiliations:

République populaire de Chine

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Le document en format XML

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<front><div type="abstract" xml:lang="en">Plant mitochondrial NAD-malic enzyme (NAD-ME), which is composed of α- and β-subunits in many species, participates in many plant biosynthetic pathways and in plant respiratory metabolism. However, little is known about the properties of woody plant NAD-MEs. In this study, we analyzed four NAD-ME genes (PtNAD-ME1 through PtNAD-ME4) in the genome of Populus trichocarpa. PtNAD-ME1 and -2 encode putative α-subunits, while PtNAD-ME3 and -4 encode putative β-subunits. The Populus NAD-MEs were expressed in Escherichia coli cells as GST-tagged fusion proteins. Each recombinant GST-PtNAD-ME protein was purified to near homogeneity by glutathione-Sepharose 4B affinity chromatography. Milligram quantities of each native protein were obtained from 1 L bacterial cultures after cleavage of the GST tag. Analysis of the enzymatic properties of these proteins in vitro indicated that α-NAD-MEs are more active than β-NAD-MEs and that α- and β-NAD-MEs presented different kinetic properties (Vmax, kcat and kcat/Km). The effect of different amounts of metabolites on the activities of Populus α- and β-NAD-MEs was assessed in vitro. While none of the metabolites evaluated in our assays activated Populus NAD-ME, oxalacetate and citrate inhibited all α- and β-NAD-MEs and glucose-6-P and fructose inhibited only the α-NAD-MEs. </div>
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Enzymatic properties of Populus α- and β-NAD-ME recombinant proteins.

Enzymatic properties of Populus α- and β-NAD-ME recombinant proteins.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki