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Glutaredoxin-dependent peroxiredoxin from poplar: protein-protein interaction and catalytic mechanism.

Identifieur interne : 004580 ( Main/Exploration ); précédent : 004579; suivant : 004581

Glutaredoxin-dependent peroxiredoxin from poplar: protein-protein interaction and catalytic mechanism.

Auteurs : Nicolas Rouhier [France] ; Eric Gelhaye ; Jean Pierre Jacquot

Source :

RBID : pubmed:11832487

Descripteurs français

English descriptors

Abstract

Recently, a poplar phloem peroxiredoxin (Prx) was found to accept both glutaredoxin (Grx) and thioredoxin (Trx) as proton donors. To investigate the catalytic mechanism of the Grx-dependent reduction of hydroperoxides catalyzed by Prx, a series of cysteinic mutants was constructed. Mutation of the most N-terminal conserved cysteine of Prx (Cys-51) demonstrates that it is the catalytic one. The second cysteine (Cys-76) is not essential for peroxiredoxin activity because the C76A mutant retained approximately 25% of the wild type Prx activity. Only one cysteine of the Grx active site (Cys-27) is essential for peroxiredoxin catalysis, indicating that Grx can act in this reaction either via a dithiol or a monothiol pathway. The creation of covalent heterodimers between Prx and Grx mutants confirms that Prx Cys-51 and Grx Cys-27 are the two residues involved in the catalytic mechanism. The integration of a third cysteine in position 152 of the Prx, making it similar in sequence to the Trx-dependent human Prx V, resulted in a protein that had no detectable activity with Grx but kept activity with Trx. Based on these experimental results, a catalytic mechanism is proposed to explain the Grx- and Trx-dependent activities of poplar Prx.

DOI: 10.1074/jbc.M111489200
PubMed: 11832487


Affiliations:


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Le document en format XML

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<term>Catalytic Domain (MeSH)</term>
<term>Cloning, Molecular (MeSH)</term>
<term>Cysteine (chemistry)</term>
<term>DNA, Complementary (metabolism)</term>
<term>Dimerization (MeSH)</term>
<term>Dose-Response Relationship, Drug (MeSH)</term>
<term>Electrophoresis, Polyacrylamide Gel (MeSH)</term>
<term>Glutaredoxins (MeSH)</term>
<term>Humans (MeSH)</term>
<term>Hydrogen Peroxide (chemistry)</term>
<term>Kinetics (MeSH)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Mutagenesis, Site-Directed (MeSH)</term>
<term>Mutation (MeSH)</term>
<term>Oxidoreductases (MeSH)</term>
<term>Oxygen (metabolism)</term>
<term>Peroxidases (genetics)</term>
<term>Peroxidases (metabolism)</term>
<term>Peroxiredoxins (MeSH)</term>
<term>Protein Binding (MeSH)</term>
<term>Protein Structure, Tertiary (MeSH)</term>
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<term>Proteins (metabolism)</term>
<term>Recombinant Proteins (metabolism)</term>
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<term>Catalyse (MeSH)</term>
<term>Cinétique (MeSH)</term>
<term>Clonage moléculaire (MeSH)</term>
<term>Cystéine (composition chimique)</term>
<term>Dimérisation (MeSH)</term>
<term>Domaine catalytique (MeSH)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Glutarédoxines (MeSH)</term>
<term>Humains (MeSH)</term>
<term>Liaison aux protéines (MeSH)</term>
<term>Mutagenèse dirigée (MeSH)</term>
<term>Mutation (MeSH)</term>
<term>Oxidoreductases (MeSH)</term>
<term>Oxygène (métabolisme)</term>
<term>Peroxidases (génétique)</term>
<term>Peroxidases (métabolisme)</term>
<term>Peroxirédoxines (MeSH)</term>
<term>Peroxyde d'hydrogène (composition chimique)</term>
<term>Protéines (génétique)</term>
<term>Protéines (métabolisme)</term>
<term>Protéines recombinantes (métabolisme)</term>
<term>Relation dose-effet des médicaments (MeSH)</term>
<term>Similitude de séquences d'acides aminés (MeSH)</term>
<term>Structure tertiaire des protéines (MeSH)</term>
<term>Séquence d'acides aminés (MeSH)</term>
<term>Technique de Western (MeSH)</term>
<term>Thiols (composition chimique)</term>
<term>Électrophorèse sur gel de polyacrylamide (MeSH)</term>
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<term>Recombinant Proteins</term>
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<term>Peroxidases</term>
<term>Protéines</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr">
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<term>Oxygène</term>
<term>Peroxidases</term>
<term>Protéines</term>
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<term>Electrophoresis, Polyacrylamide Gel</term>
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<term>Humans</term>
<term>Kinetics</term>
<term>Molecular Sequence Data</term>
<term>Mutagenesis, Site-Directed</term>
<term>Mutation</term>
<term>Oxidoreductases</term>
<term>Peroxiredoxins</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
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<term>Domaine catalytique</term>
<term>Données de séquences moléculaires</term>
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<div type="abstract" xml:lang="en">Recently, a poplar phloem peroxiredoxin (Prx) was found to accept both glutaredoxin (Grx) and thioredoxin (Trx) as proton donors. To investigate the catalytic mechanism of the Grx-dependent reduction of hydroperoxides catalyzed by Prx, a series of cysteinic mutants was constructed. Mutation of the most N-terminal conserved cysteine of Prx (Cys-51) demonstrates that it is the catalytic one. The second cysteine (Cys-76) is not essential for peroxiredoxin activity because the C76A mutant retained approximately 25% of the wild type Prx activity. Only one cysteine of the Grx active site (Cys-27) is essential for peroxiredoxin catalysis, indicating that Grx can act in this reaction either via a dithiol or a monothiol pathway. The creation of covalent heterodimers between Prx and Grx mutants confirms that Prx Cys-51 and Grx Cys-27 are the two residues involved in the catalytic mechanism. The integration of a third cysteine in position 152 of the Prx, making it similar in sequence to the Trx-dependent human Prx V, resulted in a protein that had no detectable activity with Grx but kept activity with Trx. Based on these experimental results, a catalytic mechanism is proposed to explain the Grx- and Trx-dependent activities of poplar Prx.</div>
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<AbstractText>Recently, a poplar phloem peroxiredoxin (Prx) was found to accept both glutaredoxin (Grx) and thioredoxin (Trx) as proton donors. To investigate the catalytic mechanism of the Grx-dependent reduction of hydroperoxides catalyzed by Prx, a series of cysteinic mutants was constructed. Mutation of the most N-terminal conserved cysteine of Prx (Cys-51) demonstrates that it is the catalytic one. The second cysteine (Cys-76) is not essential for peroxiredoxin activity because the C76A mutant retained approximately 25% of the wild type Prx activity. Only one cysteine of the Grx active site (Cys-27) is essential for peroxiredoxin catalysis, indicating that Grx can act in this reaction either via a dithiol or a monothiol pathway. The creation of covalent heterodimers between Prx and Grx mutants confirms that Prx Cys-51 and Grx Cys-27 are the two residues involved in the catalytic mechanism. The integration of a third cysteine in position 152 of the Prx, making it similar in sequence to the Trx-dependent human Prx V, resulted in a protein that had no detectable activity with Grx but kept activity with Trx. Based on these experimental results, a catalytic mechanism is proposed to explain the Grx- and Trx-dependent activities of poplar Prx.</AbstractText>
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HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i   -Sk "pubmed:11832487" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd   \
       | NlmPubMed2Wicri -a PoplarV1 

Wicri

This area was generated with Dilib version V0.6.37.
Data generation: Wed Nov 18 12:07:19 2020. Site generation: Wed Nov 18 12:16:31 2020