Autoprocessing mechanism of severe acute respiratory syndrome coronavirus 3C‐like protease (SARS‐CoV 3CLpro) from its polyproteins
Identifieur interne : 002626 ( Ncbi/Checkpoint ); précédent : 002625; suivant : 002627Autoprocessing mechanism of severe acute respiratory syndrome coronavirus 3C‐like protease (SARS‐CoV 3CLpro) from its polyproteins
Auteurs : Tomonari Muramatsu ; Yong-Tae Kim ; Wataru Nishii ; Takaho Terada ; Mikako Shirouzu ; Shigeyuki YokoyamaSource :
- The Febs Journal [ 1742-464X ] ; 2013.
Descripteurs français
- KwdFr :
- Biosynthèse des protéines, Cinétique, Cysteine endopeptidases (), Cysteine endopeptidases (biosynthèse), Cysteine endopeptidases (génétique), Dosages enzymatiques, Escherichia coli, Maturation post-traductionnelle des protéines, Mutagenèse dirigée, Polyprotéines (), Polyprotéines (biosynthèse), Polyprotéines (génétique), Protéines de fusion recombinantes (), Protéines de fusion recombinantes (biosynthèse), Protéines virales (), Protéines virales (biosynthèse), Protéines virales (génétique), Protéines à fluorescence verte (), Protéines à fluorescence verte (biosynthèse), Protéolyse, Précurseurs de protéines (), Précurseurs de protéines (biosynthèse), Précurseurs de protéines (génétique), Substitution d'acide aminé, Séquence d'acides aminés, Virus du SRAS (enzymologie).
- MESH :
- biosynthèse : Cysteine endopeptidases, Polyprotéines, Protéines de fusion recombinantes, Protéines virales, Protéines à fluorescence verte, Précurseurs de protéines.
- enzymologie : Virus du SRAS.
- génétique : Cysteine endopeptidases, Polyprotéines, Protéines virales, Précurseurs de protéines.
- Biosynthèse des protéines, Cinétique, Cysteine endopeptidases, Dosages enzymatiques, Escherichia coli, Maturation post-traductionnelle des protéines, Mutagenèse dirigée, Polyprotéines, Protéines de fusion recombinantes, Protéines virales, Protéines à fluorescence verte, Protéolyse, Précurseurs de protéines, Substitution d'acide aminé, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence, Amino Acid Substitution, Cysteine Endopeptidases (biosynthesis), Cysteine Endopeptidases (chemistry), Cysteine Endopeptidases (genetics), Enzyme Assays, Escherichia coli, Green Fluorescent Proteins (biosynthesis), Green Fluorescent Proteins (chemistry), Kinetics, Mutagenesis, Site-Directed, Polyproteins (biosynthesis), Polyproteins (chemistry), Polyproteins (genetics), Protein Biosynthesis, Protein Precursors (biosynthesis), Protein Precursors (chemistry), Protein Precursors (genetics), Protein Processing, Post-Translational, Proteolysis, Recombinant Fusion Proteins (biosynthesis), Recombinant Fusion Proteins (chemistry), SARS Virus (enzymology), Viral Proteins (biosynthesis), Viral Proteins (chemistry), Viral Proteins (genetics).
- MESH :
- chemical , biosynthesis : Cysteine Endopeptidases, Green Fluorescent Proteins, Polyproteins, Protein Precursors, Recombinant Fusion Proteins, Viral Proteins.
- chemical , chemistry : Cysteine Endopeptidases, Green Fluorescent Proteins, Polyproteins, Protein Precursors, Recombinant Fusion Proteins, Viral Proteins.
- chemical , genetics : Cysteine Endopeptidases, Polyproteins, Protein Precursors, Viral Proteins.
- enzymology : SARS Virus.
- Amino Acid Sequence, Amino Acid Substitution, Enzyme Assays, Escherichia coli, Kinetics, Mutagenesis, Site-Directed, Protein Biosynthesis, Protein Processing, Post-Translational, Proteolysis.
Abstract
Like many other
Url:
DOI: 10.1111/febs.12222
PubMed: 23452147
PubMed Central: 7164132
Affiliations:
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PMC:7164132Le document en format XML
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‐like protease (<styled-content style="fixed-case" toggle="no">SARS</styled-content>
‐<styled-content style="fixed-case" toggle="no">C</styled-content>
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<term>Cysteine Endopeptidases (chemistry)</term>
<term>Cysteine Endopeptidases (genetics)</term>
<term>Enzyme Assays</term>
<term>Escherichia coli</term>
<term>Green Fluorescent Proteins (biosynthesis)</term>
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<term>Kinetics</term>
<term>Mutagenesis, Site-Directed</term>
<term>Polyproteins (biosynthesis)</term>
<term>Polyproteins (chemistry)</term>
<term>Polyproteins (genetics)</term>
<term>Protein Biosynthesis</term>
<term>Protein Precursors (biosynthesis)</term>
<term>Protein Precursors (chemistry)</term>
<term>Protein Precursors (genetics)</term>
<term>Protein Processing, Post-Translational</term>
<term>Proteolysis</term>
<term>Recombinant Fusion Proteins (biosynthesis)</term>
<term>Recombinant Fusion Proteins (chemistry)</term>
<term>SARS Virus (enzymology)</term>
<term>Viral Proteins (biosynthesis)</term>
<term>Viral Proteins (chemistry)</term>
<term>Viral Proteins (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Biosynthèse des protéines</term>
<term>Cinétique</term>
<term>Cysteine endopeptidases ()</term>
<term>Cysteine endopeptidases (biosynthèse)</term>
<term>Cysteine endopeptidases (génétique)</term>
<term>Dosages enzymatiques</term>
<term>Escherichia coli</term>
<term>Maturation post-traductionnelle des protéines</term>
<term>Mutagenèse dirigée</term>
<term>Polyprotéines ()</term>
<term>Polyprotéines (biosynthèse)</term>
<term>Polyprotéines (génétique)</term>
<term>Protéines de fusion recombinantes ()</term>
<term>Protéines de fusion recombinantes (biosynthèse)</term>
<term>Protéines virales ()</term>
<term>Protéines virales (biosynthèse)</term>
<term>Protéines virales (génétique)</term>
<term>Protéines à fluorescence verte ()</term>
<term>Protéines à fluorescence verte (biosynthèse)</term>
<term>Protéolyse</term>
<term>Précurseurs de protéines ()</term>
<term>Précurseurs de protéines (biosynthèse)</term>
<term>Précurseurs de protéines (génétique)</term>
<term>Substitution d'acide aminé</term>
<term>Séquence d'acides aminés</term>
<term>Virus du SRAS (enzymologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Cysteine Endopeptidases</term>
<term>Green Fluorescent Proteins</term>
<term>Polyproteins</term>
<term>Protein Precursors</term>
<term>Recombinant Fusion Proteins</term>
<term>Viral Proteins</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Cysteine Endopeptidases</term>
<term>Green Fluorescent Proteins</term>
<term>Polyproteins</term>
<term>Protein Precursors</term>
<term>Recombinant Fusion Proteins</term>
<term>Viral Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Cysteine Endopeptidases</term>
<term>Polyproteins</term>
<term>Protein Precursors</term>
<term>Viral Proteins</term>
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<term>Polyprotéines</term>
<term>Protéines de fusion recombinantes</term>
<term>Protéines virales</term>
<term>Protéines à fluorescence verte</term>
<term>Précurseurs de protéines</term>
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<term>Protéines virales</term>
<term>Précurseurs de protéines</term>
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<keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Amino Acid Substitution</term>
<term>Enzyme Assays</term>
<term>Escherichia coli</term>
<term>Kinetics</term>
<term>Mutagenesis, Site-Directed</term>
<term>Protein Biosynthesis</term>
<term>Protein Processing, Post-Translational</term>
<term>Proteolysis</term>
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<term>Cinétique</term>
<term>Cysteine endopeptidases</term>
<term>Dosages enzymatiques</term>
<term>Escherichia coli</term>
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<term>Protéines virales</term>
<term>Protéines à fluorescence verte</term>
<term>Protéolyse</term>
<term>Précurseurs de protéines</term>
<term>Substitution d'acide aminé</term>
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<front><div type="abstract" xml:lang="en"><p>Like many other <styled-content style="fixed-case" toggle="no">RNA</styled-content>
viruses, severe acute respiratory syndrome coronavirus (<styled-content style="fixed-case" toggle="no">SARS</styled-content>
‐<styled-content style="fixed-case" toggle="no">C</styled-content>
o<styled-content style="fixed-case" toggle="no">V</styled-content>
) produces polyproteins containing several non‐structural proteins, which are then processed by the viral proteases. These proteases often exist within the polyproteins, and are excised by their own proteolytic activity (‘autoprocessing’). It is important to investigate the autoprocessing mechanism of these proteases from the point of view of anti‐<styled-content style="fixed-case" toggle="no">SARS</styled-content>
‐<styled-content style="fixed-case" toggle="no">C</styled-content>
o<styled-content style="fixed-case" toggle="no">V</styled-content>
drug design. In this paper, we describe a new method for investigating the autoprocessing mechanism of the main protease (<styled-content style="fixed-case" toggle="no">M</styled-content>
<sup>pro</sup>
), which is also called the 3<styled-content style="fixed-case" toggle="no">C</styled-content>
‐like protease (3<styled-content style="fixed-case" toggle="no">CL</styled-content>
<sup>pro</sup>
). Using our method, we measured the activities, under the same conditions, of the mature form and pro‐forms with the <styled-content style="fixed-case" toggle="no">N</styled-content>
‐terminal pro‐sequence, the <styled-content style="fixed-case" toggle="no">C</styled-content>
‐terminal pro‐sequence or both pro‐sequences, toward the pro‐form with both <styled-content style="fixed-case" toggle="no">N</styled-content>
‐ and <styled-content style="fixed-case" toggle="no">C</styled-content>
‐terminal pro‐sequences. The data indicate that the pro‐forms of the enzyme have proteolytic activity, and are stimulated by the same proteolytic activity. The stimulation occurs in two steps, with approximately eightfold stimulation by <styled-content style="fixed-case" toggle="no">N</styled-content>
‐terminal cleavage, approximately fourfold stimulation by <styled-content style="fixed-case" toggle="no">C</styled-content>
‐terminal cleavage, and 23‐fold stimulation by the cleavage of both termini, compared to the pro‐form with both the <styled-content style="fixed-case" toggle="no">N</styled-content>
‐ and <styled-content style="fixed-case" toggle="no">C</styled-content>
‐terminal pro‐sequences. Such cleavage mainly occurs in a <italic>trans</italic>
manner; i.e. the pro‐form dimer cleaves the monomeric form. The stimulation by <styled-content style="fixed-case" toggle="no">N</styled-content>
‐terminal pro‐sequence removal is due to the <italic>cis</italic>
(intra‐dimer and inter‐protomer) effect of formation of the new <styled-content style="fixed-case" toggle="no">N</styled-content>
‐terminus, whereas that by <styled-content style="fixed-case" toggle="no">C</styled-content>
‐terminal cleavage is due to removal of its <italic>trans</italic>
(inter‐dimer) inhibitory effect. A numerical simulation of the maturation pathway is presented.</p>
</div>
</front>
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<tree><noCountry><name sortKey="Kim, Yong Ae" sort="Kim, Yong Ae" uniqKey="Kim Y" first="Yong-Tae" last="Kim">Yong-Tae Kim</name>
<name sortKey="Muramatsu, Tomonari" sort="Muramatsu, Tomonari" uniqKey="Muramatsu T" first="Tomonari" last="Muramatsu">Tomonari Muramatsu</name>
<name sortKey="Nishii, Wataru" sort="Nishii, Wataru" uniqKey="Nishii W" first="Wataru" last="Nishii">Wataru Nishii</name>
<name sortKey="Shirouzu, Mikako" sort="Shirouzu, Mikako" uniqKey="Shirouzu M" first="Mikako" last="Shirouzu">Mikako Shirouzu</name>
<name sortKey="Terada, Takaho" sort="Terada, Takaho" uniqKey="Terada T" first="Takaho" last="Terada">Takaho Terada</name>
<name sortKey="Yokoyama, Shigeyuki" sort="Yokoyama, Shigeyuki" uniqKey="Yokoyama S" first="Shigeyuki" last="Yokoyama">Shigeyuki Yokoyama</name>
</noCountry>
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