SrasV1, Main, Exploration, bibRecord, 005425

Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.

Identifieur interne : 005425 ( Main/Exploration ); précédent : 005424; suivant : 005426

Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.

Auteurs : Cheng-Wen Lin [République populaire de Chine] ; Chang-Hai Tsai ; Fuu-Jen Tsai ; Pei-Jer Chen ; Chien-Chen Lai ; Lei Wan ; Hua-Hao Chiu ; Kuan-Hsun Lin

Source :

FEBS letters [ 0014-5793 ] ; 2004.

RBID : pubmed:15358553

Descripteurs français

KwdFr :
- Amorces ADN, Animaux, Antiviraux (pharmacologie), Cellules Vero, Cysteine endopeptidases, Endopeptidases (métabolisme), Hydrolyse, Protéines virales (métabolisme), Séquence nucléotidique, Virus du SRAS (), Virus du SRAS (enzymologie).
MESH :
- enzymologie : Virus du SRAS.
- métabolisme : Endopeptidases, Protéines virales.
- pharmacologie : Antiviraux.
- Amorces ADN, Animaux, Cellules Vero, Cysteine endopeptidases, Hydrolyse, Séquence nucléotidique, Virus du SRAS.

English descriptors

KwdEn :
- Animals, Antiviral Agents (pharmacology), Base Sequence, Chlorocebus aethiops, Cysteine Endopeptidases, DNA Primers, Endopeptidases (metabolism), Hydrolysis, SARS Virus (drug effects), SARS Virus (enzymology), Vero Cells, Viral Proteins (metabolism).
MESH :
- chemical , metabolism : Endopeptidases, Viral Proteins.
- chemical , pharmacology : Antiviral Agents.
- drug effects : SARS Virus.
- enzymology : SARS Virus.
- Animals, Base Sequence, Chlorocebus aethiops, Cysteine Endopeptidases, DNA Primers, Hydrolysis, Vero Cells.

Abstract

Severe acute respiratory syndrome (SARS) has been globally reported. A novel coronavirus (CoV), SARS-CoV, was identified as the etiological agent of the disease. SARS-CoV 3C-like protease (3CLpro) mediates the proteolytic processing of replicase polypeptides 1a and 1ab into functional proteins, playing an important role in viral replication. In this study, we demonstrated the expression of the SARS-CoV 3CLpro in Escherichia coli and Vero cells, and then characterized the in vitro trans-cleavage and the cell-based cis-cleavage by the 3CLpro. Mutational analysis of the 3CLpro demonstrated the importance of His41, Cys145, and Glu166 in the substrate-binding subsite S1 for keeping the proteolytic activity. In addition, alanine substitution of the cleavage substrates indicated that Gln-(P1) in the substrates mainly determined the cleavage efficiency. Therefore, this study not only established the quantifiable and reliable assay for the in vitro and cell-based measurement of the 3CLpro activity, but also characterized the molecular interaction of the SARS-CoV 3CLpro with the substrates. The results will be useful for the rational development of the anti-SARS drugs.

DOI: 10.1016/j.febslet.2004.08.017
PubMed: 15358553

Affiliations:

République populaire de Chine

Le document en format XML

<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.</title>
<author><name sortKey="Lin, Cheng Wen" sort="Lin, Cheng Wen" uniqKey="Lin C" first="Cheng-Wen" last="Lin">Cheng-Wen Lin</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung 404, Taiwan, ROC. cwlin@mail.cmu.edu.tw</nlm:affiliation>
<country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country>
<wicri:regionArea>Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung 404, Taiwan</wicri:regionArea>
<wicri:noRegion>Taiwan</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Tsai, Chang Hai" sort="Tsai, Chang Hai" uniqKey="Tsai C" first="Chang-Hai" last="Tsai">Chang-Hai Tsai</name>
</author>
<author><name sortKey="Tsai, Fuu Jen" sort="Tsai, Fuu Jen" uniqKey="Tsai F" first="Fuu-Jen" last="Tsai">Fuu-Jen Tsai</name>
</author>
<author><name sortKey="Chen, Pei Jer" sort="Chen, Pei Jer" uniqKey="Chen P" first="Pei-Jer" last="Chen">Pei-Jer Chen</name>
</author>
<author><name sortKey="Lai, Chien Chen" sort="Lai, Chien Chen" uniqKey="Lai C" first="Chien-Chen" last="Lai">Chien-Chen Lai</name>
</author>
<author><name sortKey="Wan, Lei" sort="Wan, Lei" uniqKey="Wan L" first="Lei" last="Wan">Lei Wan</name>
</author>
<author><name sortKey="Chiu, Hua Hao" sort="Chiu, Hua Hao" uniqKey="Chiu H" first="Hua-Hao" last="Chiu">Hua-Hao Chiu</name>
</author>
<author><name sortKey="Lin, Kuan Hsun" sort="Lin, Kuan Hsun" uniqKey="Lin K" first="Kuan-Hsun" last="Lin">Kuan-Hsun Lin</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2004">2004</date>
<idno type="RBID">pubmed:15358553</idno>
<idno type="pmid">15358553</idno>
<idno type="doi">10.1016/j.febslet.2004.08.017</idno>
<idno type="wicri:Area/PubMed/Corpus">002B59</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002B59</idno>
<idno type="wicri:Area/PubMed/Curation">002B59</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002B59</idno>
<idno type="wicri:Area/PubMed/Checkpoint">002D90</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002D90</idno>
<idno type="wicri:Area/Ncbi/Merge">000A68</idno>
<idno type="wicri:Area/Ncbi/Curation">000A68</idno>
<idno type="wicri:Area/Ncbi/Checkpoint">000A68</idno>
<idno type="wicri:doubleKey">0014-5793:2004:Lin C:characterization:of:trans</idno>
<idno type="wicri:Area/Main/Merge">005863</idno>
<idno type="wicri:Area/Main/Curation">005425</idno>
<idno type="wicri:Area/Main/Exploration">005425</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.</title>
<author><name sortKey="Lin, Cheng Wen" sort="Lin, Cheng Wen" uniqKey="Lin C" first="Cheng-Wen" last="Lin">Cheng-Wen Lin</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung 404, Taiwan, ROC. cwlin@mail.cmu.edu.tw</nlm:affiliation>
<country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country>
<wicri:regionArea>Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung 404, Taiwan</wicri:regionArea>
<wicri:noRegion>Taiwan</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Tsai, Chang Hai" sort="Tsai, Chang Hai" uniqKey="Tsai C" first="Chang-Hai" last="Tsai">Chang-Hai Tsai</name>
</author>
<author><name sortKey="Tsai, Fuu Jen" sort="Tsai, Fuu Jen" uniqKey="Tsai F" first="Fuu-Jen" last="Tsai">Fuu-Jen Tsai</name>
</author>
<author><name sortKey="Chen, Pei Jer" sort="Chen, Pei Jer" uniqKey="Chen P" first="Pei-Jer" last="Chen">Pei-Jer Chen</name>
</author>
<author><name sortKey="Lai, Chien Chen" sort="Lai, Chien Chen" uniqKey="Lai C" first="Chien-Chen" last="Lai">Chien-Chen Lai</name>
</author>
<author><name sortKey="Wan, Lei" sort="Wan, Lei" uniqKey="Wan L" first="Lei" last="Wan">Lei Wan</name>
</author>
<author><name sortKey="Chiu, Hua Hao" sort="Chiu, Hua Hao" uniqKey="Chiu H" first="Hua-Hao" last="Chiu">Hua-Hao Chiu</name>
</author>
<author><name sortKey="Lin, Kuan Hsun" sort="Lin, Kuan Hsun" uniqKey="Lin K" first="Kuan-Hsun" last="Lin">Kuan-Hsun Lin</name>
</author>
</analytic>
<series><title level="j">FEBS letters</title>
<idno type="ISSN">0014-5793</idno>
<imprint><date when="2004" type="published">2004</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Antiviral Agents (pharmacology)</term>
<term>Base Sequence</term>
<term>Chlorocebus aethiops</term>
<term>Cysteine Endopeptidases</term>
<term>DNA Primers</term>
<term>Endopeptidases (metabolism)</term>
<term>Hydrolysis</term>
<term>SARS Virus (drug effects)</term>
<term>SARS Virus (enzymology)</term>
<term>Vero Cells</term>
<term>Viral Proteins (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Amorces ADN</term>
<term>Animaux</term>
<term>Antiviraux (pharmacologie)</term>
<term>Cellules Vero</term>
<term>Cysteine endopeptidases</term>
<term>Endopeptidases (métabolisme)</term>
<term>Hydrolyse</term>
<term>Protéines virales (métabolisme)</term>
<term>Séquence nucléotidique</term>
<term>Virus du SRAS ()</term>
<term>Virus du SRAS (enzymologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Endopeptidases</term>
<term>Viral Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antiviral Agents</term>
</keywords>
<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Endopeptidases</term>
<term>Protéines virales</term>
</keywords>
<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Antiviraux</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Chlorocebus aethiops</term>
<term>Cysteine Endopeptidases</term>
<term>DNA Primers</term>
<term>Hydrolysis</term>
<term>Vero Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Amorces ADN</term>
<term>Animaux</term>
<term>Cellules Vero</term>
<term>Cysteine endopeptidases</term>
<term>Hydrolyse</term>
<term>Séquence nucléotidique</term>
<term>Virus du SRAS</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) has been globally reported. A novel coronavirus (CoV), SARS-CoV, was identified as the etiological agent of the disease. SARS-CoV 3C-like protease (3CLpro) mediates the proteolytic processing of replicase polypeptides 1a and 1ab into functional proteins, playing an important role in viral replication. In this study, we demonstrated the expression of the SARS-CoV 3CLpro in Escherichia coli and Vero cells, and then characterized the in vitro trans-cleavage and the cell-based cis-cleavage by the 3CLpro. Mutational analysis of the 3CLpro demonstrated the importance of His41, Cys145, and Glu166 in the substrate-binding subsite S1 for keeping the proteolytic activity. In addition, alanine substitution of the cleavage substrates indicated that Gln-(P1) in the substrates mainly determined the cleavage efficiency. Therefore, this study not only established the quantifiable and reliable assay for the in vitro and cell-based measurement of the 3CLpro activity, but also characterized the molecular interaction of the SARS-CoV 3CLpro with the substrates. The results will be useful for the rational development of the anti-SARS drugs.</div>
</front>
</TEI>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
</list>
<tree><noCountry><name sortKey="Chen, Pei Jer" sort="Chen, Pei Jer" uniqKey="Chen P" first="Pei-Jer" last="Chen">Pei-Jer Chen</name>
<name sortKey="Chiu, Hua Hao" sort="Chiu, Hua Hao" uniqKey="Chiu H" first="Hua-Hao" last="Chiu">Hua-Hao Chiu</name>
<name sortKey="Lai, Chien Chen" sort="Lai, Chien Chen" uniqKey="Lai C" first="Chien-Chen" last="Lai">Chien-Chen Lai</name>
<name sortKey="Lin, Kuan Hsun" sort="Lin, Kuan Hsun" uniqKey="Lin K" first="Kuan-Hsun" last="Lin">Kuan-Hsun Lin</name>
<name sortKey="Tsai, Chang Hai" sort="Tsai, Chang Hai" uniqKey="Tsai C" first="Chang-Hai" last="Tsai">Chang-Hai Tsai</name>
<name sortKey="Tsai, Fuu Jen" sort="Tsai, Fuu Jen" uniqKey="Tsai F" first="Fuu-Jen" last="Tsai">Fuu-Jen Tsai</name>
<name sortKey="Wan, Lei" sort="Wan, Lei" uniqKey="Wan L" first="Lei" last="Wan">Lei Wan</name>
</noCountry>
<country name="République populaire de Chine"><noRegion><name sortKey="Lin, Cheng Wen" sort="Lin, Cheng Wen" uniqKey="Lin C" first="Cheng-Wen" last="Lin">Cheng-Wen Lin</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>

Pour manipuler ce document sous Unix (Dilib)

EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration

HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 005425 | SxmlIndent | more

HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 005425 | SxmlIndent | more

Pour mettre un lien sur cette page dans le réseau Wicri

{{Explor lien
   |wiki=    Sante
   |area=    SrasV1
   |flux=    Main
   |étape=   Exploration
   |type=    RBID
   |clé=     pubmed:15358553
   |texte=   Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.
}}

Pour générer des pages wiki

HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i   -Sk "pubmed:15358553" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd   \
       | NlmPubMed2Wicri -a SrasV1

This area was generated with Dilib version V0.6.33.
Data generation: Tue Apr 28 14:49:16 2020. Site generation: Sat Mar 27 22:06:49 2021

Serveur d'exploration SRAS

Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.

Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

	Serveur d'exploration SRAS
	Attention, ce site est en cours de développement ! Attention, site généré par des moyens informatiques à partir de corpus bruts. Les informations ne sont donc pas validées.