Crystallization and preliminary X-ray diffraction analysis of Nsp15 from SARS coronavirus.
Identifieur interne : 000307 ( France/Analysis ); précédent : 000306; suivant : 000308Crystallization and preliminary X-ray diffraction analysis of Nsp15 from SARS coronavirus.
Auteurs : Stéfano Ricagno [France] ; Bruno Coutard ; Sacha Grisel ; Nicolas Brémond ; Karen Dalle ; Fabienne Tocque ; Valérie Campanacci ; Julie Lichière ; Violaine Lantez ; Claire Debarnot ; Christian Cambillau ; Bruno Canard ; Marie Pierre EgloffSource :
- Acta crystallographica. Section F, Structural biology and crystallization communications [ 1744-3091 ] ; 2006.
Descripteurs français
- KwdFr :
- Clonage moléculaire, Conformation des protéines, Cristallisation, Diffraction des rayons X, Escherichia coli, Fragments peptidiques (), Protéines recombinantes (), Protéines recombinantes (isolement et purification), Protéines virales non structurales (), Protéines virales non structurales (génétique), Protéines virales non structurales (isolement et purification), RNA replicase (), RNA replicase (génétique), RNA replicase (isolement et purification), Structures macromoléculaires (), Structures macromoléculaires (isolement et purification), Virus du SRAS ().
- MESH :
- génétique : Protéines virales non structurales, RNA replicase.
- isolement et purification : Protéines recombinantes, Protéines virales non structurales, RNA replicase, Structures macromoléculaires.
- Clonage moléculaire, Conformation des protéines, Cristallisation, Diffraction des rayons X, Escherichia coli, Fragments peptidiques, Protéines recombinantes, Protéines virales non structurales, RNA replicase, Structures macromoléculaires, Virus du SRAS.
English descriptors
- KwdEn :
- Cloning, Molecular, Crystallization, Escherichia coli, Macromolecular Substances (chemistry), Macromolecular Substances (isolation & purification), Peptide Fragments (chemistry), Protein Conformation, RNA Replicase (chemistry), RNA Replicase (genetics), RNA Replicase (isolation & purification), Recombinant Proteins (chemistry), Recombinant Proteins (isolation & purification), SARS Virus (chemistry), Viral Nonstructural Proteins (chemistry), Viral Nonstructural Proteins (genetics), Viral Nonstructural Proteins (isolation & purification), X-Ray Diffraction.
- MESH :
- chemical , chemistry : Macromolecular Substances, Peptide Fragments, RNA Replicase, Recombinant Proteins, Viral Nonstructural Proteins.
- chemical , genetics : RNA Replicase, Viral Nonstructural Proteins.
- chemical , isolation & purification : Macromolecular Substances, RNA Replicase, Recombinant Proteins, Viral Nonstructural Proteins.
- chemistry : SARS Virus.
- Cloning, Molecular, Crystallization, Escherichia coli, Protein Conformation, X-Ray Diffraction.
Abstract
The non-structural protein Nsp15 from the aetiological agent of SARS (severe acute respiratory syndrome) has recently been characterized as a uridine-specific endoribonuclease. This enzyme plays an essential role in viral replication and transcription since a mutation in the related H229E human coronavirus nsp15 gene can abolish viral RNA synthesis. SARS full-length Nsp15 (346 amino acids) has been cloned and expressed in Escherichia coli with an N-terminal hexahistidine tag and has been purified to homogeneity. The protein was subsequently crystallized using PEG 8000 or 10 000 as precipitants. Small cubic crystals of the apoenzyme were obtained from 100 nl nanodrops. They belong to space group P4(1)32 or P4(3)32, with unit-cell parameters a = b = c = 166.8 angstroms. Diffraction data were collected to a maximum resolution of 2.7 angstroms.
DOI: 10.1107/S1744309106009407
PubMed: 16582498
Affiliations:
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analysis of Nsp15 from SARS coronavirus.</title><author><name sortKey="Ricagno, Stefano" sort="Ricagno, Stefano" uniqKey="Ricagno S" first="Stéfano" last="Ricagno">Stéfano Ricagno</name><affiliation wicri:level="3"><nlm:affiliation>Centre National de la Recherche Scientifique and Universités d'Aix-Marseille I et II, UMR 6098, Architecture et Fonction des Macromolécules Biologiques, Ecole Supérieure d'Ingénieurs de Luminy-Case 925, 13288 Marseille CEDEX 9, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Centre National de la Recherche Scientifique and Universités d'Aix-Marseille I et II, UMR 6098, Architecture et Fonction des Macromolécules Biologiques, Ecole Supérieure d'Ingénieurs de Luminy-Case 925, 13288 Marseille CEDEX 9</wicri:regionArea><placeName><region type="region" nuts="2">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName></affiliation></author><author><name sortKey="Coutard, Bruno" sort="Coutard, 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This enzyme plays an essential role in viral replication and transcription since a mutation in the related H229E human coronavirus nsp15 gene can abolish viral RNA synthesis. SARS full-length Nsp15 (346 amino acids) has been cloned and expressed in Escherichia coli with an N-terminal hexahistidine tag and has been purified to homogeneity. The protein was subsequently crystallized using PEG 8000 or 10 000 as precipitants. Small cubic crystals of the apoenzyme were obtained from 100 nl nanodrops. They belong to space group P4(1)32 or P4(3)32, with unit-cell parameters a = b = c = 166.8 angstroms. Diffraction data were collected to a maximum resolution of 2.7 angstroms.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Provence-Alpes-Côte d'Azur</li></region><settlement><li>Marseille</li></settlement></list><tree><noCountry><name sortKey="Bremond, Nicolas" sort="Bremond, Nicolas" uniqKey="Bremond N" first="Nicolas" last="Brémond">Nicolas Brémond</name><name sortKey="Cambillau, Christian" sort="Cambillau, Christian" uniqKey="Cambillau C" first="Christian" last="Cambillau">Christian Cambillau</name><name sortKey="Campanacci, Valerie" sort="Campanacci, Valerie" uniqKey="Campanacci V" first="Valérie" last="Campanacci">Valérie Campanacci</name><name sortKey="Canard, Bruno" sort="Canard, Bruno" uniqKey="Canard B" first="Bruno" last="Canard">Bruno Canard</name><name sortKey="Coutard, Bruno" sort="Coutard, Bruno" uniqKey="Coutard B" first="Bruno" last="Coutard">Bruno Coutard</name><name sortKey="Dalle, Karen" sort="Dalle, Karen" uniqKey="Dalle K" first="Karen" last="Dalle">Karen Dalle</name><name sortKey="Debarnot, Claire" sort="Debarnot, Claire" uniqKey="Debarnot C" first="Claire" last="Debarnot">Claire Debarnot</name><name sortKey="Egloff, Marie Pierre" sort="Egloff, Marie Pierre" uniqKey="Egloff M" first="Marie Pierre" last="Egloff">Marie Pierre Egloff</name><name sortKey="Grisel, Sacha" sort="Grisel, Sacha" uniqKey="Grisel S" first="Sacha" last="Grisel">Sacha Grisel</name><name sortKey="Lantez, Violaine" sort="Lantez, Violaine" uniqKey="Lantez V" first="Violaine" last="Lantez">Violaine Lantez</name><name sortKey="Lichiere, Julie" sort="Lichiere, Julie" uniqKey="Lichiere J" first="Julie" last="Lichière">Julie Lichière</name><name sortKey="Tocque, Fabienne" sort="Tocque, Fabienne" uniqKey="Tocque F" first="Fabienne" last="Tocque">Fabienne Tocque</name></noCountry><country name="France"><region name="Provence-Alpes-Côte d'Azur"><name sortKey="Ricagno, Stefano" sort="Ricagno, Stefano" uniqKey="Ricagno S" first="Stéfano" last="Ricagno">Stéfano Ricagno</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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