Crystallization and preliminary X-ray crystallographic analysis of a nonstructural protein 15 mutant from Human coronavirus 229E.
Identifieur interne : 001339 ( Main/Exploration ); précédent : 001338; suivant : 001340Crystallization and preliminary X-ray crystallographic analysis of a nonstructural protein 15 mutant from Human coronavirus 229E.
Auteurs : Tong Huo [République populaire de Chine] ; Xiang Liu [République populaire de Chine]Source :
- Acta crystallographica. Section F, Structural biology communications [ 2053-230X ] ; 2015.
Descripteurs français
- KwdFr :
- Chromatographie sur gel, Coronavirus humain 229E (), Cristallisation, Cristallographie aux rayons X, Données de séquences moléculaires, Masse moléculaire, Protéines mutantes (), Protéines virales non structurales (), Séquence d'acides aminés, Ultracentrifugation, Électrophorèse sur gel de polyacrylamide.
- MESH :
English descriptors
- KwdEn :
- MESH :
Abstract
Nonstructural protein 15 (nsp15), also called endoribonuclease, is a gene product of open reading frame 1b (ORF 1b) in coronaviruses. It is an important enzyme in the transcription/replication process involved in discontinuous negative-strand RNA synthesis. In this work, mutants of nsp15 from Human coronavirus 229E (HCoV-229E) were made based on structural analysis of the homologous nsp15s in Severe acute respiratory syndrome coronavirus (SARS-CoV) and Mouse hepatitis virus (MHV). The I26A/N52A mutant of nsp15 was overexpressed, purified and crystallized, and this mutant led to a trimeric form rather than hexamers or monomers. Crystals of trimeric nsp15 were obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant and diffracted to 2.5 Å resolution. The crystals belonged to space group C2221, with unit-cell parameters a = 85.9, b = 137.5, c = 423.1 Å, α = β = γ = 90°.
DOI: 10.1107/S2053230X15007359
PubMed: 26323302
Affiliations:
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Section F, Structural biology communications</title><idno type="eISSN">2053-230X</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Chromatography, Gel</term><term>Coronavirus 229E, Human (chemistry)</term><term>Crystallization</term><term>Crystallography, X-Ray</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Molecular Sequence Data</term><term>Molecular Weight</term><term>Mutant Proteins (chemistry)</term><term>Ultracentrifugation</term><term>Viral Nonstructural Proteins (chemistry)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Chromatographie sur gel</term><term>Coronavirus humain 229E ()</term><term>Cristallisation</term><term>Cristallographie aux rayons X</term><term>Données de séquences moléculaires</term><term>Masse moléculaire</term><term>Protéines mutantes ()</term><term>Protéines virales non structurales ()</term><term>Séquence d'acides aminés</term><term>Ultracentrifugation</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Mutant Proteins</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Coronavirus 229E, Human</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Chromatography, Gel</term><term>Crystallization</term><term>Crystallography, X-Ray</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Molecular Sequence Data</term><term>Molecular Weight</term><term>Ultracentrifugation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Chromatographie sur gel</term><term>Coronavirus humain 229E</term><term>Cristallisation</term><term>Cristallographie aux rayons X</term><term>Données de séquences moléculaires</term><term>Masse moléculaire</term><term>Protéines mutantes</term><term>Protéines virales non structurales</term><term>Séquence d'acides aminés</term><term>Ultracentrifugation</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Nonstructural protein 15 (nsp15), also called endoribonuclease, is a gene product of open reading frame 1b (ORF 1b) in coronaviruses. It is an important enzyme in the transcription/replication process involved in discontinuous negative-strand RNA synthesis. In this work, mutants of nsp15 from Human coronavirus 229E (HCoV-229E) were made based on structural analysis of the homologous nsp15s in Severe acute respiratory syndrome coronavirus (SARS-CoV) and Mouse hepatitis virus (MHV). The I26A/N52A mutant of nsp15 was overexpressed, purified and crystallized, and this mutant led to a trimeric form rather than hexamers or monomers. Crystals of trimeric nsp15 were obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant and diffracted to 2.5 Å resolution. The crystals belonged to space group C2221, with unit-cell parameters a = 85.9, b = 137.5, c = 423.1 Å, α = β = γ = 90°. </div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Tianjin</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Huo, Tong" sort="Huo, Tong" uniqKey="Huo T" first="Tong" last="Huo">Tong Huo</name></noRegion><name sortKey="Liu, Xiang" sort="Liu, Xiang" uniqKey="Liu X" first="Xiang" last="Liu">Xiang Liu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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