Biochemical characterization of exoribonuclease encoded by SARS coronavirus.
Identifieur interne : 003765 ( Main/Exploration ); précédent : 003764; suivant : 003766Biochemical characterization of exoribonuclease encoded by SARS coronavirus.
Auteurs : Ping Chen [République populaire de Chine] ; Miao Jiang ; Tao Hu ; Qingzhen Liu ; Xiaojiang S. Chen ; Deyin GuoSource :
- Journal of biochemistry and molecular biology [ 1225-8687 ] ; 2007.
Descripteurs français
- KwdFr :
- Activation enzymatique (), Calorimétrie, Cations divalents (métabolisme), Cations divalents (pharmacologie), Cinétique, Dichroïsme circulaire, Escherichia coli (génétique), Exoribonucleases (), Exoribonucleases (génétique), Exoribonucleases (métabolisme), Liaison aux protéines, Magnésium (métabolisme), Magnésium (pharmacologie), Manganèse (métabolisme), Manganèse (pharmacologie), Protéines recombinantes (), Protéines recombinantes (métabolisme), Protéines virales (), Protéines virales (génétique), Protéines virales (métabolisme), Protéines virales non structurales (), Protéines virales non structurales (génétique), Protéines virales non structurales (métabolisme), Sites de fixation, Spécificité du substrat, Thermodynamique, Virus du SRAS (enzymologie), Zinc (métabolisme), Zinc (pharmacologie).
- MESH :
- enzymologie : Virus du SRAS.
- génétique : Escherichia coli, Exoribonucleases, Protéines virales, Protéines virales non structurales.
- métabolisme : Cations divalents, Exoribonucleases, Magnésium, Manganèse, Protéines recombinantes, Protéines virales, Protéines virales non structurales, Zinc.
- pharmacologie : Cations divalents, Magnésium, Manganèse, Zinc.
- Activation enzymatique, Calorimétrie, Cinétique, Dichroïsme circulaire, Exoribonucleases, Liaison aux protéines, Protéines recombinantes, Protéines virales, Protéines virales non structurales, Sites de fixation, Spécificité du substrat, Thermodynamique.
English descriptors
- KwdEn :
- Binding Sites, Calorimetry, Cations, Divalent (metabolism), Cations, Divalent (pharmacology), Circular Dichroism, Enzyme Activation (drug effects), Escherichia coli (genetics), Exoribonucleases (chemistry), Exoribonucleases (genetics), Exoribonucleases (metabolism), Kinetics, Magnesium (metabolism), Magnesium (pharmacology), Manganese (metabolism), Manganese (pharmacology), Protein Binding, Recombinant Proteins (chemistry), Recombinant Proteins (metabolism), SARS Virus (enzymology), Substrate Specificity, Thermodynamics, Viral Nonstructural Proteins (chemistry), Viral Nonstructural Proteins (genetics), Viral Nonstructural Proteins (metabolism), Viral Proteins (chemistry), Viral Proteins (genetics), Viral Proteins (metabolism), Zinc (metabolism), Zinc (pharmacology).
- MESH :
- chemical , chemistry : Exoribonucleases, Recombinant Proteins, Viral Nonstructural Proteins, Viral Proteins.
- chemical , genetics : Exoribonucleases, Viral Nonstructural Proteins, Viral Proteins.
- chemical , metabolism : Cations, Divalent, Exoribonucleases, Magnesium, Manganese, Recombinant Proteins, Viral Nonstructural Proteins, Viral Proteins, Zinc.
- chemical , pharmacology : Cations, Divalent, Magnesium, Manganese, Zinc.
- drug effects : Enzyme Activation.
- enzymology : SARS Virus.
- genetics : Escherichia coli.
- Binding Sites, Calorimetry, Circular Dichroism, Kinetics, Protein Binding, Substrate Specificity, Thermodynamics.
Abstract
The nsp14 protein is an exoribonuclease that is encoded by severe acute respiratory syndrome coronavirus (SARS-CoV). We have cloned and expressed the nsp14 protein in Escherichia coli, and characterized the nature and the role(s) of the metal ions in the reaction chemistry. The purified recombinant nsp14 protein digested a 5'-labeled RNA molecule, but failed to digest the RNA substrate that is modified with fluorescein group at the 3'-hydroxyl group, suggesting a 3'-to-5' exoribonuclease activity. The exoribonuclease activity requires Mg2+ as a cofactor. Isothermal titration calorimetry (ITC) analysis indicated a two-metal binding mode for divalent cations by nsp14. Endogenous tryptophan fluorescence and circular dichroism (CD) spectra measurements showed that there was a structural change of nsp14 when binding with metal ions. We propose that the conformational change induced by metal ions may be a prerequisite for catalytic activity by correctly positioning the side chains of the residues located in the active site of the enzyme.
DOI: 10.5483/bmbrep.2007.40.5.649
PubMed: 17927896
Affiliations:
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Le document en format XML
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Chen</name></author><author><name sortKey="Guo, Deyin" sort="Guo, Deyin" uniqKey="Guo D" first="Deyin" last="Guo">Deyin Guo</name></author></analytic><series><title level="j">Journal of biochemistry and molecular biology</title><idno type="ISSN">1225-8687</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Binding Sites</term><term>Calorimetry</term><term>Cations, Divalent (metabolism)</term><term>Cations, Divalent (pharmacology)</term><term>Circular Dichroism</term><term>Enzyme Activation (drug effects)</term><term>Escherichia coli (genetics)</term><term>Exoribonucleases (chemistry)</term><term>Exoribonucleases (genetics)</term><term>Exoribonucleases (metabolism)</term><term>Kinetics</term><term>Magnesium (metabolism)</term><term>Magnesium (pharmacology)</term><term>Manganese (metabolism)</term><term>Manganese (pharmacology)</term><term>Protein Binding</term><term>Recombinant Proteins (chemistry)</term><term>Recombinant Proteins (metabolism)</term><term>SARS Virus (enzymology)</term><term>Substrate Specificity</term><term>Thermodynamics</term><term>Viral Nonstructural Proteins (chemistry)</term><term>Viral Nonstructural Proteins (genetics)</term><term>Viral Nonstructural Proteins (metabolism)</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (metabolism)</term><term>Zinc (metabolism)</term><term>Zinc (pharmacology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Activation enzymatique ()</term><term>Calorimétrie</term><term>Cations divalents (métabolisme)</term><term>Cations divalents (pharmacologie)</term><term>Cinétique</term><term>Dichroïsme circulaire</term><term>Escherichia coli (génétique)</term><term>Exoribonucleases ()</term><term>Exoribonucleases (génétique)</term><term>Exoribonucleases (métabolisme)</term><term>Liaison aux protéines</term><term>Magnésium (métabolisme)</term><term>Magnésium (pharmacologie)</term><term>Manganèse (métabolisme)</term><term>Manganèse (pharmacologie)</term><term>Protéines recombinantes ()</term><term>Protéines recombinantes (métabolisme)</term><term>Protéines virales ()</term><term>Protéines virales (génétique)</term><term>Protéines virales (métabolisme)</term><term>Protéines virales non structurales ()</term><term>Protéines virales non structurales (génétique)</term><term>Protéines virales non structurales (métabolisme)</term><term>Sites de fixation</term><term>Spécificité du substrat</term><term>Thermodynamique</term><term>Virus du SRAS (enzymologie)</term><term>Zinc (métabolisme)</term><term>Zinc (pharmacologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Exoribonucleases</term><term>Recombinant Proteins</term><term>Viral Nonstructural Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" 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xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Escherichia coli</term><term>Exoribonucleases</term><term>Protéines virales</term><term>Protéines virales non structurales</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cations divalents</term><term>Exoribonucleases</term><term>Magnésium</term><term>Manganèse</term><term>Protéines recombinantes</term><term>Protéines virales</term><term>Protéines virales non structurales</term><term>Zinc</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Cations divalents</term><term>Magnésium</term><term>Manganèse</term><term>Zinc</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Binding Sites</term><term>Calorimetry</term><term>Circular Dichroism</term><term>Kinetics</term><term>Protein Binding</term><term>Substrate Specificity</term><term>Thermodynamics</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Activation enzymatique</term><term>Calorimétrie</term><term>Cinétique</term><term>Dichroïsme circulaire</term><term>Exoribonucleases</term><term>Liaison aux protéines</term><term>Protéines recombinantes</term><term>Protéines virales</term><term>Protéines virales non structurales</term><term>Sites de fixation</term><term>Spécificité du substrat</term><term>Thermodynamique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The nsp14 protein is an exoribonuclease that is encoded by severe acute respiratory syndrome coronavirus (SARS-CoV). We have cloned and expressed the nsp14 protein in Escherichia coli, and characterized the nature and the role(s) of the metal ions in the reaction chemistry. The purified recombinant nsp14 protein digested a 5'-labeled RNA molecule, but failed to digest the RNA substrate that is modified with fluorescein group at the 3'-hydroxyl group, suggesting a 3'-to-5' exoribonuclease activity. The exoribonuclease activity requires Mg2+ as a cofactor. Isothermal titration calorimetry (ITC) analysis indicated a two-metal binding mode for divalent cations by nsp14. Endogenous tryptophan fluorescence and circular dichroism (CD) spectra measurements showed that there was a structural change of nsp14 when binding with metal ions. We propose that the conformational change induced by metal ions may be a prerequisite for catalytic activity by correctly positioning the side chains of the residues located in the active site of the enzyme.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Hubei</li></region><settlement><li>Wuhan</li></settlement><orgName><li>Université de Wuhan</li></orgName></list><tree><noCountry><name sortKey="Chen, Xiaojiang S" sort="Chen, Xiaojiang S" uniqKey="Chen X" first="Xiaojiang S" last="Chen">Xiaojiang S. Chen</name><name sortKey="Guo, Deyin" sort="Guo, Deyin" uniqKey="Guo D" first="Deyin" last="Guo">Deyin Guo</name><name sortKey="Hu, Tao" sort="Hu, Tao" uniqKey="Hu T" first="Tao" last="Hu">Tao Hu</name><name sortKey="Jiang, Miao" sort="Jiang, Miao" uniqKey="Jiang M" first="Miao" last="Jiang">Miao Jiang</name><name sortKey="Liu, Qingzhen" sort="Liu, Qingzhen" uniqKey="Liu Q" first="Qingzhen" last="Liu">Qingzhen Liu</name></noCountry><country name="République populaire de Chine"><region name="Hubei"><name sortKey="Chen, Ping" sort="Chen, Ping" uniqKey="Chen P" first="Ping" last="Chen">Ping Chen</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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