Substrate specificity profiling and identification of a new class of inhibitor for the major protease of the SARS coronavirus.
Identifieur interne : 003584 ( Main/Exploration ); précédent : 003583; suivant : 003585Substrate specificity profiling and identification of a new class of inhibitor for the major protease of the SARS coronavirus.
Auteurs : D H Goetz [États-Unis] ; Y. Choe ; E. Hansell ; Y T Chen ; M. Mcdowell ; C B Jonsson ; W R Roush ; J. Mckerrow ; C S CraikSource :
- Biochemistry [ 0006-2960 ] ; 2007.
Descripteurs français
- KwdFr :
- Animaux, Antiviraux (), Antiviraux (isolement et purification), Antiviraux (métabolisme), Antiviraux (pharmacologie), Banque de peptides, Cellules Vero, Composés époxy (), Composés époxy (isolement et purification), Composés époxy (métabolisme), Composés époxy (pharmacologie), Cristallographie aux rayons X, Cysteine endopeptidases (), Dipeptides (), Dipeptides (isolement et purification), Dipeptides (métabolisme), Dipeptides (pharmacologie), Domaine catalytique (), Inhibiteurs de la cystéine protéinase (), Inhibiteurs de la cystéine protéinase (pharmacologie), Modèles moléculaires, Oligopeptides (), Oligopeptides (isolement et purification), Oligopeptides (métabolisme), Oligopeptides (pharmacologie), Protéines virales (), Protéines virales (antagonistes et inhibiteurs), Relation structure-activité, Réplication virale (), Spécificité du substrat, Structure tertiaire des protéines, Substitution d'acide aminé, Virus du SRAS (), Virus du SRAS (enzymologie).
- MESH :
- antagonistes et inhibiteurs : Protéines virales.
- enzymologie : Virus du SRAS.
- isolement et purification : Antiviraux, Composés époxy, Dipeptides, Oligopeptides.
- métabolisme : Antiviraux, Composés époxy, Dipeptides, Oligopeptides.
- pharmacologie : Antiviraux, Composés époxy, Dipeptides, Inhibiteurs de la cystéine protéinase, Oligopeptides.
- Animaux, Antiviraux, Banque de peptides, Cellules Vero, Composés époxy, Cristallographie aux rayons X, Cysteine endopeptidases, Dipeptides, Domaine catalytique, Inhibiteurs de la cystéine protéinase, Modèles moléculaires, Oligopeptides, Protéines virales, Relation structure-activité, Réplication virale, Spécificité du substrat, Structure tertiaire des protéines, Substitution d'acide aminé, Virus du SRAS.
English descriptors
- KwdEn :
- Amino Acid Substitution, Animals, Antiviral Agents (chemistry), Antiviral Agents (isolation & purification), Antiviral Agents (metabolism), Antiviral Agents (pharmacology), Catalytic Domain (drug effects), Chlorocebus aethiops, Crystallography, X-Ray, Cysteine Endopeptidases (chemistry), Cysteine Proteinase Inhibitors (chemistry), Cysteine Proteinase Inhibitors (classification), Cysteine Proteinase Inhibitors (pharmacology), Dipeptides (chemistry), Dipeptides (isolation & purification), Dipeptides (metabolism), Dipeptides (pharmacology), Epoxy Compounds (chemistry), Epoxy Compounds (isolation & purification), Epoxy Compounds (metabolism), Epoxy Compounds (pharmacology), Models, Molecular, Oligopeptides (chemistry), Oligopeptides (isolation & purification), Oligopeptides (metabolism), Oligopeptides (pharmacology), Peptide Library, Protein Structure, Tertiary, SARS Virus (drug effects), SARS Virus (enzymology), Structure-Activity Relationship, Substrate Specificity, Vero Cells, Viral Proteins (antagonists & inhibitors), Viral Proteins (chemistry), Virus Replication (drug effects).
- MESH :
- chemical , antagonists & inhibitors : Viral Proteins.
- chemical , chemistry : Antiviral Agents, Cysteine Endopeptidases, Cysteine Proteinase Inhibitors, Dipeptides, Epoxy Compounds, Oligopeptides, Viral Proteins.
- chemical , classification : Cysteine Proteinase Inhibitors.
- chemical , isolation & purification : Antiviral Agents, Dipeptides, Epoxy Compounds, Oligopeptides.
- chemical , metabolism : Antiviral Agents, Dipeptides, Epoxy Compounds, Oligopeptides.
- chemical , pharmacology : Antiviral Agents, Cysteine Proteinase Inhibitors, Dipeptides, Epoxy Compounds, Oligopeptides.
- drug effects : Catalytic Domain, SARS Virus, Virus Replication.
- enzymology : SARS Virus.
- Amino Acid Substitution, Animals, Chlorocebus aethiops, Crystallography, X-Ray, Models, Molecular, Peptide Library, Protein Structure, Tertiary, Structure-Activity Relationship, Substrate Specificity, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a high rate of mortality. The SARS-associated coronavirus (SARS-CoV) has been identified as the etiological agent of the disease. Although public health procedures have been effective in combating the spread of SARS, concern remains about the possibility of a recurrence. Various approaches are being pursued for the development of efficacious therapeutics. One promising approach is to develop small molecule inhibitors of the essential major polyprotein processing protease 3Clpro. Here we report a complete description of the tetrapeptide substrate specificity of 3Clpro using fully degenerate peptide libraries consisting of all 160,000 possible naturally occurring tetrapeptides. The substrate specificity data show the expected P1-Gln P2-Leu specificity and elucidate a novel preference for P1-His containing substrates equal to the expected preference for P1-Gln. These data were then used to develop optimal substrates for a high-throughput screen of a 2000 compound small-molecule inhibitor library consisting of known cysteine protease inhibitor scaffolds. We also report the 1.8 A X-ray crystal structure of 3Clpro bound to an irreversible inhibitor. This inhibitor, an alpha,beta-epoxyketone, inhibits 3Clpro with a k3/Ki of 0.002 microM(-1) s(-1) in a mode consistent with the substrate specificity data. Finally, we report the successful rational improvement of this scaffold with second generation inhibitors. These data provide the foundation for a rational small-molecule inhibitor design effort based upon the inhibitor scaffold identified, the crystal structure of the complex, and a more complete understanding of P1-P4 substrate specificity.
DOI: 10.1021/bi0621415
PubMed: 17605471
Affiliations:
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Hansell</name></author><author><name sortKey="Chen, Y T" sort="Chen, Y T" uniqKey="Chen Y" first="Y T" last="Chen">Y T Chen</name></author><author><name sortKey="Mcdowell, M" sort="Mcdowell, M" uniqKey="Mcdowell M" first="M" last="Mcdowell">M. Mcdowell</name></author><author><name sortKey="Jonsson, C B" sort="Jonsson, C B" uniqKey="Jonsson C" first="C B" last="Jonsson">C B Jonsson</name></author><author><name sortKey="Roush, W R" sort="Roush, W R" uniqKey="Roush W" first="W R" last="Roush">W R Roush</name></author><author><name sortKey="Mckerrow, J" sort="Mckerrow, J" uniqKey="Mckerrow J" first="J" last="Mckerrow">J. 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Mckerrow</name></author><author><name sortKey="Craik, C S" sort="Craik, C S" uniqKey="Craik C" first="C S" last="Craik">C S Craik</name></author></analytic><series><title level="j">Biochemistry</title><idno type="ISSN">0006-2960</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Substitution</term><term>Animals</term><term>Antiviral Agents (chemistry)</term><term>Antiviral Agents (isolation & purification)</term><term>Antiviral Agents (metabolism)</term><term>Antiviral Agents (pharmacology)</term><term>Catalytic Domain (drug effects)</term><term>Chlorocebus aethiops</term><term>Crystallography, X-Ray</term><term>Cysteine Endopeptidases (chemistry)</term><term>Cysteine Proteinase Inhibitors (chemistry)</term><term>Cysteine Proteinase Inhibitors (classification)</term><term>Cysteine Proteinase Inhibitors (pharmacology)</term><term>Dipeptides 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xml:lang="fr"><term>Animaux</term><term>Antiviraux</term><term>Banque de peptides</term><term>Cellules Vero</term><term>Composés époxy</term><term>Cristallographie aux rayons X</term><term>Cysteine endopeptidases</term><term>Dipeptides</term><term>Domaine catalytique</term><term>Inhibiteurs de la cystéine protéinase</term><term>Modèles moléculaires</term><term>Oligopeptides</term><term>Protéines virales</term><term>Relation structure-activité</term><term>Réplication virale</term><term>Spécificité du substrat</term><term>Structure tertiaire des protéines</term><term>Substitution d'acide aminé</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a high rate of mortality. The SARS-associated coronavirus (SARS-CoV) has been identified as the etiological agent of the disease. Although public health procedures have been effective in combating the spread of SARS, concern remains about the possibility of a recurrence. Various approaches are being pursued for the development of efficacious therapeutics. One promising approach is to develop small molecule inhibitors of the essential major polyprotein processing protease 3Clpro. Here we report a complete description of the tetrapeptide substrate specificity of 3Clpro using fully degenerate peptide libraries consisting of all 160,000 possible naturally occurring tetrapeptides. The substrate specificity data show the expected P1-Gln P2-Leu specificity and elucidate a novel preference for P1-His containing substrates equal to the expected preference for P1-Gln. These data were then used to develop optimal substrates for a high-throughput screen of a 2000 compound small-molecule inhibitor library consisting of known cysteine protease inhibitor scaffolds. We also report the 1.8 A X-ray crystal structure of 3Clpro bound to an irreversible inhibitor. This inhibitor, an alpha,beta-epoxyketone, inhibits 3Clpro with a k3/Ki of 0.002 microM(-1) s(-1) in a mode consistent with the substrate specificity data. Finally, we report the successful rational improvement of this scaffold with second generation inhibitors. These data provide the foundation for a rational small-molecule inhibitor design effort based upon the inhibitor scaffold identified, the crystal structure of the complex, and a more complete understanding of P1-P4 substrate specificity.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Chen, Y T" sort="Chen, Y T" uniqKey="Chen Y" first="Y T" last="Chen">Y T Chen</name><name sortKey="Choe, Y" sort="Choe, Y" uniqKey="Choe Y" first="Y" last="Choe">Y. Choe</name><name sortKey="Craik, C S" sort="Craik, C S" uniqKey="Craik C" first="C S" last="Craik">C S Craik</name><name sortKey="Hansell, E" sort="Hansell, E" uniqKey="Hansell E" first="E" last="Hansell">E. Hansell</name><name sortKey="Jonsson, C B" sort="Jonsson, C B" uniqKey="Jonsson C" first="C B" last="Jonsson">C B Jonsson</name><name sortKey="Mcdowell, M" sort="Mcdowell, M" uniqKey="Mcdowell M" first="M" last="Mcdowell">M. Mcdowell</name><name sortKey="Mckerrow, J" sort="Mckerrow, J" uniqKey="Mckerrow J" first="J" last="Mckerrow">J. Mckerrow</name><name sortKey="Roush, W R" sort="Roush, W R" uniqKey="Roush W" first="W R" last="Roush">W R Roush</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Goetz, D H" sort="Goetz, D H" uniqKey="Goetz D" first="D H" last="Goetz">D H Goetz</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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