Structure of Main Protease from Human Coronavirus NL63: Insights for Wide Spectrum Anti-Coronavirus Drug Design
Identifieur interne : 000064 ( Pmc/Curation ); précédent : 000063; suivant : 000065Structure of Main Protease from Human Coronavirus NL63: Insights for Wide Spectrum Anti-Coronavirus Drug Design
Auteurs : Fenghua Wang [République populaire de Chine] ; Cheng Chen [République populaire de Chine] ; Wenjie Tan [République populaire de Chine] ; Kailin Yang [États-Unis] ; Haitao Yang [République populaire de Chine]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
First identified in The Netherlands in 2004, human coronavirus NL63 (HCoV-NL63) was found to cause worldwide infections. Patients infected by HCoV-NL63 are typically young children with upper and lower respiratory tract infection, presenting with symptoms including croup, bronchiolitis, and pneumonia. Unfortunately, there are currently no effective antiviral therapy to contain HCoV-NL63 infection. CoV genomes encode an integral viral component, main protease (Mpro), which is essential for viral replication through proteolytic processing of RNA replicase machinery. Due to the sequence and structural conservation among all CoVs, Mpro has been recognized as an attractive molecular target for rational anti-CoV drug design. Here we present the crystal structure of HCoV-NL63 Mpro in complex with a Michael acceptor inhibitor N3. Structural analysis, consistent with biochemical inhibition results, reveals the molecular mechanism of enzyme inhibition at the highly conservative substrate-recognition pocket. We show such molecular target remains unchanged across 30 clinical isolates of HCoV-NL63 strains. Through comparative study with Mpros from other human CoVs (including the deadly SARS-CoV and MERS-CoV) and their related zoonotic CoVs, our structure of HCoV-NL63 Mpro provides critical insight into rational development of wide spectrum antiviral therapeutics to treat infections caused by human CoVs.
Url:
DOI: 10.1038/srep22677
PubMed: 26948040
PubMed Central: 4780191
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country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Tianjin International Joint Academy of Biotechnology and Medicine</institution>, Tianjin 300457,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention</institution>, Beijing 102206,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Kailin" sort="Yang, Kailin" uniqKey="Yang K" first="Kailin" last="Yang">Kailin Yang</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Cleveland Clinic Lerner College of Medicine of Case Western Reserve University</institution>, Cleveland, OH 44195,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Haitao" sort="Yang, Haitao" uniqKey="Yang H" first="Haitao" last="Yang">Haitao Yang</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>School of Life Sciences, Tianjin University</institution>, Tianjin 300072,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Tianjin International Joint Academy of Biotechnology and Medicine</institution>, Tianjin 300457,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26948040</idno><idno type="pmc">4780191</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4780191</idno><idno type="RBID">PMC:4780191</idno><idno type="doi">10.1038/srep22677</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000064</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000064</idno><idno type="wicri:Area/Pmc/Curation">000064</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000064</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Structure of Main Protease from Human Coronavirus NL63: Insights for Wide Spectrum Anti-Coronavirus Drug Design</title><author><name sortKey="Wang, Fenghua" sort="Wang, Fenghua" uniqKey="Wang F" first="Fenghua" last="Wang">Fenghua Wang</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>School of Life Sciences, Tianjin University</institution>, Tianjin 300072,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Chen, Cheng" sort="Chen, Cheng" uniqKey="Chen C" first="Cheng" last="Chen">Cheng Chen</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>School of Life Sciences, Tianjin University</institution>, Tianjin 300072,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Tianjin International Joint Academy of Biotechnology and Medicine</institution>, Tianjin 300457,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention</institution>, Beijing 102206,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Kailin" sort="Yang, Kailin" uniqKey="Yang K" first="Kailin" last="Yang">Kailin Yang</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Cleveland Clinic Lerner College of Medicine of Case Western Reserve University</institution>, Cleveland, OH 44195,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Haitao" sort="Yang, Haitao" uniqKey="Yang H" first="Haitao" last="Yang">Haitao Yang</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>School of Life Sciences, Tianjin University</institution>, Tianjin 300072,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Tianjin International Joint Academy of Biotechnology and Medicine</institution>, Tianjin 300457,<country>China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>First identified in The Netherlands in 2004, human coronavirus NL63 (HCoV-NL63) was found to cause worldwide infections. Patients infected by HCoV-NL63 are typically young children with upper and lower respiratory tract infection, presenting with symptoms including croup, bronchiolitis, and pneumonia. Unfortunately, there are currently no effective antiviral therapy to contain HCoV-NL63 infection. CoV genomes encode an integral viral component, main protease (M<sup>pro</sup>), which is essential for viral replication through proteolytic processing of RNA replicase machinery. Due to the sequence and structural conservation among all CoVs, M<sup>pro</sup> has been recognized as an attractive molecular target for rational anti-CoV drug design. Here we present the crystal structure of HCoV-NL63 M<sup>pro</sup> in complex with a Michael acceptor inhibitor N3. Structural analysis, consistent with biochemical inhibition results, reveals the molecular mechanism of enzyme inhibition at the highly conservative substrate-recognition pocket. We show such molecular target remains unchanged across 30 clinical isolates of HCoV-NL63 strains. Through comparative study with M<sup>pro</sup>s from other human CoVs (including the deadly SARS-CoV and MERS-CoV) and their related zoonotic CoVs, our structure of HCoV-NL63 M<sup>pro</sup> provides critical insight into rational development of wide spectrum antiviral therapeutics to treat infections caused by human CoVs.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Weiss, S R" uniqKey="Weiss S">S. R. Weiss</name></author><author><name sortKey="Navas Martin, S" uniqKey="Navas Martin S">S. Navas-Martin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Masters, P S" uniqKey="Masters P">P. S. Masters</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Siddell, S" uniqKey="Siddell S">S. Siddell</name></author><author><name sortKey="Wege, H" uniqKey="Wege H">H. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26948040</article-id><article-id pub-id-type="pmc">4780191</article-id><article-id pub-id-type="pii">srep22677</article-id><article-id pub-id-type="doi">10.1038/srep22677</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Structure of Main Protease from Human Coronavirus NL63: Insights for Wide Spectrum Anti-Coronavirus Drug Design</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Fenghua</given-names></name><xref ref-type="aff" rid="a1">1</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Chen</surname><given-names>Cheng</given-names></name><xref ref-type="aff" rid="a1">1</xref><xref ref-type="aff" rid="a2">2</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Tan</surname><given-names>Wenjie</given-names></name><xref ref-type="aff" rid="a3">3</xref></contrib><contrib contrib-type="author"><name><surname>Yang</surname><given-names>Kailin</given-names></name><xref ref-type="corresp" rid="c1">a</xref><xref ref-type="aff" rid="a4">4</xref></contrib><contrib contrib-type="author"><name><surname>Yang</surname><given-names>Haitao</given-names></name><xref ref-type="corresp" rid="c2">b</xref><xref ref-type="aff" rid="a1">1</xref><xref ref-type="aff" rid="a2">2</xref></contrib><aff id="a1"><label>1</label><institution>School of Life Sciences, Tianjin University</institution>, Tianjin 300072,<country>China</country></aff><aff id="a2"><label>2</label><institution>Tianjin International Joint Academy of Biotechnology and Medicine</institution>, Tianjin 300457,<country>China</country></aff><aff id="a3"><label>3</label><institution>Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention</institution>, Beijing 102206,<country>China</country></aff><aff id="a4"><label>4</label><institution>Cleveland Clinic Lerner College of Medicine of Case Western Reserve University</institution>, Cleveland, OH 44195,<country>USA</country></aff></contrib-group><author-notes><corresp id="c1"><label>a</label><email>yangk2@ccf.org</email></corresp><corresp id="c2"><label>b</label><email>yanght@tju.edu.cn</email></corresp><fn id="n1"><label>*</label><p>These authors contributed equally to this work.</p></fn></author-notes><pub-date pub-type="epub"><day>07</day><month>03</month><year>2016</year></pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><volume>6</volume><elocation-id>22677</elocation-id><history><date date-type="received"><day>04</day><month>12</month><year>2015</year></date><date date-type="accepted"><day>17</day><month>02</month><year>2016</year></date></history><permissions><copyright-statement>Copyright © 2016, Macmillan Publishers Limited</copyright-statement><copyright-year>2016</copyright-year><copyright-holder>Macmillan Publishers Limited</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link></license-p></license></permissions><abstract><p>First identified in The Netherlands in 2004, human coronavirus NL63 (HCoV-NL63) was found to cause worldwide infections. Patients infected by HCoV-NL63 are typically young children with upper and lower respiratory tract infection, presenting with symptoms including croup, bronchiolitis, and pneumonia. Unfortunately, there are currently no effective antiviral therapy to contain HCoV-NL63 infection. CoV genomes encode an integral viral component, main protease (M<sup>pro</sup>), which is essential for viral replication through proteolytic processing of RNA replicase machinery. Due to the sequence and structural conservation among all CoVs, M<sup>pro</sup> has been recognized as an attractive molecular target for rational anti-CoV drug design. Here we present the crystal structure of HCoV-NL63 M<sup>pro</sup> in complex with a Michael acceptor inhibitor N3. Structural analysis, consistent with biochemical inhibition results, reveals the molecular mechanism of enzyme inhibition at the highly conservative substrate-recognition pocket. We show such molecular target remains unchanged across 30 clinical isolates of HCoV-NL63 strains. Through comparative study with M<sup>pro</sup>s from other human CoVs (including the deadly SARS-CoV and MERS-CoV) and their related zoonotic CoVs, our structure of HCoV-NL63 M<sup>pro</sup> provides critical insight into rational development of wide spectrum antiviral therapeutics to treat infections caused by human CoVs.</p></abstract></article-meta></front><floats-group><fig id="f1"><label>Figure 1</label><caption><title>Structural overview of HCoV-NL63 M<sup>pro</sup>.</title><p>(<bold>a</bold>) Overview of homodimer in one asymmetric unit (A: slate and B: deep salmon). Protomers are shown in cartoons, and N3 inhibitors are shown as green sticks. (<bold>b</bold>) Structural alignment of protomer A in M<sup>pro</sup>-N3 (slate) complex with that in apo enzyme (light orange, PDB ID: 3TLO; C. P. Chuck & K. B. Wong, unpublished work). The backbone is shown in cartoons, and N3 inhibitor is presented as green sticks.</p></caption><graphic xlink:href="srep22677-f1"></graphic></fig><fig id="f2"><label>Figure 2</label><caption><title>Structure of inhibitor N3 and its interaction with HCoV-NL63 M<sup>pro</sup>.</title><p>(<bold>a)</bold> The structure of compound N3. (<bold>b</bold>) A stereo view of N3 bound to the substrate-binding pocket of HCoV-NL63 M<sup>pro</sup> at 2.85 Å. N3 inhibitor is shown in green and covered by an omit map contoured at 1.0 σ. Residues forming the substrate-binding pocket are shown in silver. <bold>(c)</bold> Detailed view of the interaction between N3 and HCoV-NL63 M<sup>pro</sup>. The N3 inhibitor is shown in green. Hydrogen bonds are shown as dashed lines labeled with interaction distances. The covalent bond between N3 and Sγ of Cys144 is labeled in red.</p></caption><graphic xlink:href="srep22677-f2"></graphic></fig><fig id="f3"><label>Figure 3</label><caption><title>Surface representation of native HCoV-NL63 and its complex with inhibitor N3.</title><p>(<bold>a)</bold> Surface representation of substrate-binding pockets from apo enzyme of HCoV-NL63 M<sup>pro</sup> (marine, PDB ID: 3TLO). The S1, S2, S4, and S1’ pockets and residues forming the substrate-binding pocket are labeled. <bold>(b)</bold> Surface representation of HCoV-NL63 M<sup>pro</sup> (marine) in complex with N3 inhibitor (green). Water molecules are shown as red spheres. The P1-P5, and P1’ groups of N3 inhibitor are labeled together with residues forming the substrate-binding pocket.</p></caption><graphic xlink:href="srep22677-f3"></graphic></fig><fig id="f4"><label>Figure 4</label><caption><title>S1 and S2 binding sites in HCoV-NL63 M<sup>pro</sup>.</title><p>The main chains of four human CoV M<sup>pro</sup> structures (HCoV-NL63: slate, HCoV-229E: cyan, SARS-CoV: magenta, and HCoV-HKU1: yellow) are superimposed and displayed in the neightborhood of the substrate-binding site. The S1, S2, and S4 binding sites are labeled. The backbones are represented in worm form, and inhibitor is shown in stick format of green color. Residues 45–51 are marked with a red oval (in dash line).</p></caption><graphic xlink:href="srep22677-f4"></graphic></fig><fig id="f5"><label>Figure 5</label><caption><title>Structural conservation of M<sup>pro</sup> among clinical HCoV-NL63 isolates.</title><p>(<bold>a</bold>) Summary of the 3 residues of variation among a total of 30 HCoV-NL63 strains. Amsterdam I is used as reference strain. Number of strains carrying one particular residue is labeled in bracket. (<bold>b</bold>) Three-dimensional representation of the 3 nonconserved residues from (<bold>a</bold>) mapped onto HCoV-NL63 M<sup>pro</sup> complex structure with inhibitor N3. Conserved residues are colored light orange, the 3 residues (His69, Cys221, and Met235) are colored cyan, and inhibitor N3 is colored green.</p></caption><graphic xlink:href="srep22677-f5"></graphic></fig><fig id="f6"><label>Figure 6</label><caption><title>Inhibitor N3 targets a conserved site among human and related zoonotic CoVs.</title><p>(<bold>a</bold>) Sequence alignment of three pairs of human CoVs and their related zoonotic counterparts: HCoV-229E and bovine coronavirus (BCoV), SARS-CoV and SARS-related coronavirus isolated from bat (BtSARSr-CoV), MERS-CoV and dromedary camel MERS-CoV (DcMERS-CoV). HCoV-NL63 and its secondary structure are used as reference for the alignment. Sequence alignment was performed using ClustalW2<xref ref-type="bibr" rid="b72">72</xref>, and figure was generated using ESPript 3.0<xref ref-type="bibr" rid="b73">73</xref>. (<bold>b</bold>) Surface representation of conserved substrate-binding pockets from 7 CoV M<sup>pro</sup>s listed in (a). Background is HCoV-NL63 M<sup>pro</sup>. Red: identical residues among all seven CoV M<sup>pro</sup>; magenta: substitution in one CoV M<sup>pro</sup>; orange: substitution in two CoV M<sup>pro</sup>s. The residues forming the substrate-binding pocket are labeled. S1’: Leu27, Gly142; S1: Phe139, His163, Glu166, His172; S2: His41 Tyr53, Asp187; S4: Leu167, Gln192.</p></caption><graphic xlink:href="srep22677-f6"></graphic></fig><table-wrap position="float" id="t1"><label>Table 1</label><caption><title>X-ray data-processing and refinement statistics.</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th align="left" valign="top" charoff="50">Statistics</th><th align="center" valign="top" charoff="50">Value for the HCoV-NL63 M<sup>pro</sup>-N3 complex</th></tr></thead><tbody valign="top"><tr><td colspan="2" align="left" valign="top" charoff="50">Data collection</td></tr><tr><td align="left" valign="top" charoff="50"> Wavelength (Å)</td><td align="center" valign="top" charoff="50">1.0000</td></tr><tr><td align="left" valign="top" charoff="50"> Resolution limit (Å)</td><td align="center" valign="top" charoff="50">50.0–2.85 (2.90–2.85)</td></tr><tr><td align="left" valign="top" charoff="50"> Space group</td><td align="center" valign="top" charoff="50"><italic>P</italic>4<sub>1</sub>2<sub>1</sub>2</td></tr><tr><td colspan="2" align="left" valign="top" charoff="50">Cell parameters</td></tr><tr><td align="left" valign="top" charoff="50"> a (Å)</td><td align="center" valign="top" charoff="50">87.2</td></tr><tr><td align="left" valign="top" charoff="50"> b (Å)</td><td align="center" valign="top" charoff="50">87.2</td></tr><tr><td align="left" valign="top" charoff="50"> c (Å)</td><td align="center" valign="top" charoff="50">212.1</td></tr><tr><td align="left" valign="top" charoff="50"> α = β = γ (°)</td><td align="center" valign="top" charoff="50">90</td></tr><tr><td align="left" valign="top" charoff="50">Total no. of reflections</td><td align="center" valign="top" charoff="50">283898</td></tr><tr><td align="left" valign="top" charoff="50">No. of unique reflections</td><td align="center" valign="top" charoff="50">19967</td></tr><tr><td align="left" valign="top" charoff="50">Completeness (%)</td><td align="center" valign="top" charoff="50">100 (100)</td></tr><tr><td align="left" valign="top" charoff="50">Redundancy</td><td align="center" valign="top" charoff="50">14.2 (12.6)</td></tr><tr><td align="left" valign="top" charoff="50">R<sub>merge</sub> (%)</td><td align="center" valign="top" charoff="50">14.4 (46.6)</td></tr><tr><td align="left" valign="top" charoff="50">Sigma cutoff</td><td align="center" valign="top" charoff="50">0</td></tr><tr><td align="left" valign="top" charoff="50"><italic>I/σ(I)</italic></td><td align="center" valign="top" charoff="50">21.5 (6.1)</td></tr><tr><td colspan="2" align="left" valign="top" charoff="50">Refinement</td></tr><tr><td align="left" valign="top" charoff="50"> Resolution range (Å)</td><td align="center" valign="top" charoff="50">50.0–2.85</td></tr><tr><td align="left" valign="top" charoff="50"> R<sub>work</sub> (%)</td><td align="center" valign="top" charoff="50">19.3</td></tr><tr><td align="left" valign="top" charoff="50"> R<sub>free</sub> (%)</td><td align="center" valign="top" charoff="50">24.1</td></tr><tr><td colspan="2" align="left" valign="top" charoff="50">Rmsd from ideal geometry</td></tr><tr><td align="left" valign="top" charoff="50"> Bonds (Å)</td><td align="center" valign="top" charoff="50">0.007</td></tr><tr><td align="left" valign="top" charoff="50"> Angles (°)</td><td align="center" valign="top" charoff="50">1.16</td></tr><tr><td align="left" valign="top" charoff="50">Avg B factor (Å<sup>2</sup>)</td><td align="center" valign="top" charoff="50">27.6</td></tr><tr><td align="left" valign="top" charoff="50"> Protein</td><td align="center" valign="top" charoff="50">28.1</td></tr><tr><td align="left" valign="top" charoff="50"> Small molecule</td><td align="center" valign="top" charoff="50">28.3</td></tr><tr><td colspan="2" align="left" valign="top" charoff="50">Ramachandran plot</td></tr><tr><td align="left" valign="top" charoff="50"> Favored (%)</td><td align="center" valign="top" charoff="50">96.0</td></tr><tr><td align="left" valign="top" charoff="50"> Allowed (%)</td><td align="center" valign="top" charoff="50">4.0</td></tr><tr><td align="left" valign="top" charoff="50"> Outliers (%)</td><td align="center" valign="top" charoff="50">0.0</td></tr></tbody></table></table-wrap><table-wrap position="float" id="t2"><label>Table 2</label><caption><title>Enzyme activity and N3 inhibition data for HCoV-NL63 M<sup>pro</sup>.</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center" char="±"></col><col align="center" char="±"></col><col align="center" char="±"></col><col align="center" char="±"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th rowspan="2" align="left" valign="top" charoff="50">Virus M<sup>pro</sup></th><th rowspan="2" align="center" valign="top" char="±" charoff="50"><italic>K</italic><sub><italic>m</italic></sub> (μM)</th><th rowspan="2" align="center" valign="top" char="±" charoff="50"><italic>k</italic><sub><italic>cat</italic></sub> (s<sup>−1</sup>)</th><th colspan="2" align="center" valign="top" char="±" charoff="50">Inhibitor N3<hr></hr></th><th rowspan="2" align="center" valign="top" charoff="50">Data Source</th></tr><tr><th align="center" valign="top" char="±" charoff="50"><italic>K</italic><sub><italic>i</italic></sub> (μM)</th><th align="center" valign="top" char="±" charoff="50"><italic>k</italic><sub><italic>3</italic></sub> (10<sup>−3</sup>∙s<sup>−1</sup>)</th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">HCoV-NL63</td><td align="center" valign="top" char="±" charoff="50">50.8 ± 3.4</td><td align="center" valign="top" char="±" charoff="50">0.098 ± 0.004</td><td align="center" valign="top" char="±" charoff="50">11.3 ± 1.0</td><td align="center" valign="top" char="±" charoff="50">42.4 ± 5.0</td><td align="center" valign="top" charoff="50">This study</td></tr><tr><td align="left" valign="top" charoff="50">SARS-CoV</td><td align="center" valign="top" char="±" charoff="50">129 ± 7</td><td align="center" valign="top" char="±" charoff="50">0.14 ± 0.01</td><td align="center" valign="top" char="±" charoff="50">9.0 ± 0.8</td><td align="center" valign="top" char="±" charoff="50">3.1 ± 0.5</td><td align="center" valign="top" charoff="50"><xref ref-type="bibr" rid="b49">49</xref></td></tr><tr><td align="left" valign="top" charoff="50">HCoV-229E</td><td align="center" valign="top" char="±" charoff="50">29.8 ± 0.9</td><td align="center" valign="top" char="±" charoff="50">1.27 ± 0.09</td><td align="center" valign="top" char="±" charoff="50">1.67 ± 0.18</td><td align="center" valign="top" char="±" charoff="50">18.0 ± 1.1</td><td align="center" valign="top" charoff="50"><xref ref-type="bibr" rid="b49">49</xref></td></tr><tr><td align="left" valign="top" charoff="50">HCoV-HKU1</td><td align="center" valign="top" char="±" charoff="50">83.2 ± 13.3</td><td align="center" valign="top" char="±" charoff="50">1.1 ± 0.12</td><td align="center" valign="top" char="±" charoff="50">−</td><td align="center" valign="top" char="±" charoff="50">−</td><td align="center" valign="top" charoff="50"><xref ref-type="bibr" rid="b53">53</xref></td></tr></tbody></table></table-wrap><table-wrap position="float" id="t3"><label>Table 3</label><caption><title>P2 residues from genomes of six human CoVs.</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th rowspan="2" align="left" valign="top" charoff="50">No.</th><th rowspan="2" align="center" valign="top" charoff="50">Cleavage Site</th><th colspan="2" align="center" valign="top" charoff="50">Alphacoronavirus<hr></hr></th><th colspan="4" align="center" valign="top" charoff="50">Betacoronavirus<hr></hr></th></tr><tr><th align="center" valign="top" charoff="50">HCoV-NL63</th><th align="center" valign="top" charoff="50">HCoV-229E</th><th align="center" valign="top" charoff="50">HCoV-HKU1</th><th align="center" valign="top" charoff="50">HCoV-OC43</th><th align="center" valign="top" charoff="50">SARS-CoV</th><th align="center" valign="top" charoff="50">MERS-CoV</th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">1</td><td align="center" valign="top" charoff="50">nsp4-nsp5</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">2</td><td align="center" valign="top" charoff="50">nsp5-nsp6</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">F</td><td align="center" valign="top" charoff="50">M</td></tr><tr><td align="left" valign="top" charoff="50">3</td><td align="center" valign="top" charoff="50">nsp6-nsp7</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">I</td><td align="center" valign="top" charoff="50">F</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">M</td></tr><tr><td align="left" valign="top" charoff="50">4</td><td align="center" valign="top" charoff="50">nsp7-nsp8</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">5</td><td align="center" valign="top" charoff="50">nsp8-nsp9</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">M</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">6</td><td align="center" valign="top" charoff="50">nsp9-nsp10</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">7</td><td align="center" valign="top" charoff="50">nsp10-nsp11</td><td align="center" valign="top" charoff="50">I</td><td align="center" valign="top" charoff="50">I</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">M</td><td align="center" valign="top" charoff="50">P</td></tr><tr><td align="left" valign="top" charoff="50">8</td><td align="center" valign="top" charoff="50">nsp12-nsp13</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">M</td><td align="center" valign="top" charoff="50">M</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">9</td><td align="center" valign="top" charoff="50">nsp13-nsp14</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">V</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr><tr><td align="left" valign="top" charoff="50">10</td><td align="center" valign="top" charoff="50">nsp14-nsp15</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">V</td></tr><tr><td align="left" valign="top" charoff="50">11</td><td align="center" valign="top" charoff="50">nsp15-nsp16</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">M</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td><td align="center" valign="top" charoff="50">L</td></tr></tbody></table></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Sante/explor/CovidV1/Data/Pmc/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000064 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd -nk 000064 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Sante
|area= CovidV1
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|type= RBID
|clé= PMC:4780191
|texte= Structure of Main Protease from Human Coronavirus NL63: Insights for Wide Spectrum Anti-Coronavirus Drug Design
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Curation/RBID.i -Sk "pubmed:26948040" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a CovidV1
| This area was generated with Dilib version V0.6.33. |  |