Engineering of the H2O2-binding pocket region of a recombinant manganese peroxidase to be resistant to H2O2.
Identifieur interne : 000A44 ( Main/Exploration ); précédent : 000A43; suivant : 000A45Engineering of the H2O2-binding pocket region of a recombinant manganese peroxidase to be resistant to H2O2.
Auteurs : C. Miyazaki [Japon] ; H. TakahashiSource :
- FEBS letters [ 0014-5793 ] ; 2001.
Descripteurs français
- KwdFr :
- Conformation des protéines (MeSH), Escherichia coli (métabolisme), Liaison aux protéines (MeSH), Modèles moléculaires (MeSH), Mutagenèse dirigée (MeSH), Mutation (MeSH), Méthionine (composition chimique), Oxygène (métabolisme), Peroxidases (composition chimique), Peroxidases (métabolisme), Peroxyde d'hydrogène (métabolisme), Phanerochaete (enzymologie), Pliage des protéines (MeSH), Protéines recombinantes (composition chimique), Protéines recombinantes (métabolisme), Relation dose-effet des médicaments (MeSH), Sites de fixation (MeSH).
- MESH :
- composition chimique : Méthionine, Peroxidases, Protéines recombinantes.
- enzymologie : Phanerochaete.
- métabolisme : Escherichia coli, Oxygène, Peroxidases, Peroxyde d'hydrogène, Protéines recombinantes.
- Conformation des protéines, Liaison aux protéines, Modèles moléculaires, Mutagenèse dirigée, Mutation, Pliage des protéines, Relation dose-effet des médicaments, Sites de fixation.
English descriptors
- KwdEn :
- Binding Sites (MeSH), Dose-Response Relationship, Drug (MeSH), Escherichia coli (metabolism), Hydrogen Peroxide (metabolism), Methionine (chemistry), Models, Molecular (MeSH), Mutagenesis, Site-Directed (MeSH), Mutation (MeSH), Oxygen (metabolism), Peroxidases (chemistry), Peroxidases (metabolism), Phanerochaete (enzymology), Protein Binding (MeSH), Protein Conformation (MeSH), Protein Folding (MeSH), Recombinant Proteins (chemistry), Recombinant Proteins (metabolism).
- MESH :
- chemical , chemistry : Methionine, Peroxidases, Recombinant Proteins.
- chemical , metabolism : Hydrogen Peroxide, Oxygen, Peroxidases, Recombinant Proteins.
- enzymology : Phanerochaete.
- metabolism : Escherichia coli.
- Binding Sites, Dose-Response Relationship, Drug, Models, Molecular, Mutagenesis, Site-Directed, Mutation, Protein Binding, Protein Conformation, Protein Folding.
Abstract
The manganese peroxidase produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+), is easily inactivated by the hydrogen peroxide (H2O2) presented in the reaction. We attempted to increase H2O2 resistance by the conformational stabilization around the H2O2-binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non-oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H2O2 where the wild-type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non-bulky and non-oxidizable Ser.
DOI: 10.1016/s0014-5793(01)03127-1
PubMed: 11734216
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<affiliation wicri:level="1"><nlm:affiliation>Toyota Central R&D Labs., 41-1, Yokomichi, Aichi 480-1192, Nagakute, Japan.</nlm:affiliation>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Binding Sites (MeSH)</term>
<term>Dose-Response Relationship, Drug (MeSH)</term>
<term>Escherichia coli (metabolism)</term>
<term>Hydrogen Peroxide (metabolism)</term>
<term>Methionine (chemistry)</term>
<term>Models, Molecular (MeSH)</term>
<term>Mutagenesis, Site-Directed (MeSH)</term>
<term>Mutation (MeSH)</term>
<term>Oxygen (metabolism)</term>
<term>Peroxidases (chemistry)</term>
<term>Peroxidases (metabolism)</term>
<term>Phanerochaete (enzymology)</term>
<term>Protein Binding (MeSH)</term>
<term>Protein Conformation (MeSH)</term>
<term>Protein Folding (MeSH)</term>
<term>Recombinant Proteins (chemistry)</term>
<term>Recombinant Proteins (metabolism)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Conformation des protéines (MeSH)</term>
<term>Escherichia coli (métabolisme)</term>
<term>Liaison aux protéines (MeSH)</term>
<term>Modèles moléculaires (MeSH)</term>
<term>Mutagenèse dirigée (MeSH)</term>
<term>Mutation (MeSH)</term>
<term>Méthionine (composition chimique)</term>
<term>Oxygène (métabolisme)</term>
<term>Peroxidases (composition chimique)</term>
<term>Peroxidases (métabolisme)</term>
<term>Peroxyde d'hydrogène (métabolisme)</term>
<term>Phanerochaete (enzymologie)</term>
<term>Pliage des protéines (MeSH)</term>
<term>Protéines recombinantes (composition chimique)</term>
<term>Protéines recombinantes (métabolisme)</term>
<term>Relation dose-effet des médicaments (MeSH)</term>
<term>Sites de fixation (MeSH)</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Methionine</term>
<term>Peroxidases</term>
<term>Recombinant Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Hydrogen Peroxide</term>
<term>Oxygen</term>
<term>Peroxidases</term>
<term>Recombinant Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Méthionine</term>
<term>Peroxidases</term>
<term>Protéines recombinantes</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Phanerochaete</term>
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<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Phanerochaete</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term>
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<term>Oxygène</term>
<term>Peroxidases</term>
<term>Peroxyde d'hydrogène</term>
<term>Protéines recombinantes</term>
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<term>Dose-Response Relationship, Drug</term>
<term>Models, Molecular</term>
<term>Mutagenesis, Site-Directed</term>
<term>Mutation</term>
<term>Protein Binding</term>
<term>Protein Conformation</term>
<term>Protein Folding</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Conformation des protéines</term>
<term>Liaison aux protéines</term>
<term>Modèles moléculaires</term>
<term>Mutagenèse dirigée</term>
<term>Mutation</term>
<term>Pliage des protéines</term>
<term>Relation dose-effet des médicaments</term>
<term>Sites de fixation</term>
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<front><div type="abstract" xml:lang="en">The manganese peroxidase produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+), is easily inactivated by the hydrogen peroxide (H2O2) presented in the reaction. We attempted to increase H2O2 resistance by the conformational stabilization around the H2O2-binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non-oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H2O2 where the wild-type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non-bulky and non-oxidizable Ser.</div>
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<Abstract><AbstractText>The manganese peroxidase produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+), is easily inactivated by the hydrogen peroxide (H2O2) presented in the reaction. We attempted to increase H2O2 resistance by the conformational stabilization around the H2O2-binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non-oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H2O2 where the wild-type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non-bulky and non-oxidizable Ser.</AbstractText>
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