Low Stability of Nucleocapsid Protein in SARS Virus†
Identifieur interne : 000009 ( Istex/Corpus ); précédent : 000008; suivant : 000010Low Stability of Nucleocapsid Protein in SARS Virus†
Auteurs : Yulong Wang ; Xiaoyu Wu ; Yihua Wang ; Bing Li ; Hao Zhou ; Guiyong Yuan ; Yan Fu ; Yongzhang LuoSource :
- Biochemistry [ 0006-2960 ] ; 2004.
Abstract
The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome (SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study, the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as shown by 1H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea (with Cm of 2.77 M and m value of 2.74 kcal mol-1 M-1) or GdmCl (with Cm of 1.46 M and m value of 4.50 kcal mol-1 M-1). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to be inactivated. We propose that the low stability of N protein may be critical for the stability and function of SARS virus.
Url:
DOI: 10.1021/bi049194b
Links to Exploration step
ISTEX:096978D90918130B3334B42F12577503A0738C60Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Low Stability of Nucleocapsid Protein in SARS Virus†</title><author><name sortKey="Wang, Yulong" sort="Wang, Yulong" uniqKey="Wang Y" first="Yulong" last="Wang">Yulong Wang</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> Y.W. and X.W. contributed equally to this study.</mods:affiliation></affiliation></author><author><name sortKey="Wu, Xiaoyu" sort="Wu, Xiaoyu" uniqKey="Wu X" first="Xiaoyu" last="Wu">Xiaoyu Wu</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> Y.W. and X.W. contributed equally to this study.</mods:affiliation></affiliation></author><author><name sortKey="Wang, Yihua" sort="Wang, Yihua" uniqKey="Wang Y" first="Yihua" last="Wang">Yihua Wang</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Li, Bing" sort="Li, Bing" uniqKey="Li B" first="Bing" last="Li">Bing Li</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Zhou, Hao" sort="Zhou, Hao" uniqKey="Zhou H" first="Hao" last="Zhou">Hao Zhou</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Yuan, Guiyong" sort="Yuan, Guiyong" uniqKey="Yuan G" first="Guiyong" last="Yuan">Guiyong Yuan</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Fu, Yan" sort="Fu, Yan" uniqKey="Fu Y" first="Yan" last="Fu">Yan Fu</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Luo, Yongzhang" sort="Luo, Yongzhang" uniqKey="Luo Y" first="Yongzhang" last="Luo">Yongzhang Luo</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:096978D90918130B3334B42F12577503A0738C60</idno><date when="2004" year="2004">2004</date><idno type="doi">10.1021/bi049194b</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000009</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000009</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">Low Stability of Nucleocapsid Protein in SARS Virus<ref type="bib" target="#bi049194bAF2">†</ref></title><author><name sortKey="Wang, Yulong" sort="Wang, Yulong" uniqKey="Wang Y" first="Yulong" last="Wang">Yulong Wang</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> Y.W. and X.W. contributed equally to this study.</mods:affiliation></affiliation></author><author><name sortKey="Wu, Xiaoyu" sort="Wu, Xiaoyu" uniqKey="Wu X" first="Xiaoyu" last="Wu">Xiaoyu Wu</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> Y.W. and X.W. contributed equally to this study.</mods:affiliation></affiliation></author><author><name sortKey="Wang, Yihua" sort="Wang, Yihua" uniqKey="Wang Y" first="Yihua" last="Wang">Yihua Wang</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Li, Bing" sort="Li, Bing" uniqKey="Li B" first="Bing" last="Li">Bing Li</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Zhou, Hao" sort="Zhou, Hao" uniqKey="Zhou H" first="Hao" last="Zhou">Hao Zhou</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Yuan, Guiyong" sort="Yuan, Guiyong" uniqKey="Yuan G" first="Guiyong" last="Yuan">Guiyong Yuan</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Fu, Yan" sort="Fu, Yan" uniqKey="Fu Y" first="Yan" last="Fu">Yan Fu</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation></author><author><name sortKey="Luo, Yongzhang" sort="Luo, Yongzhang" uniqKey="Luo Y" first="Yongzhang" last="Luo">Yongzhang Luo</name><affiliation><mods:affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Biochemistry</title><title level="j" type="abbrev">Biochemistry</title><idno type="ISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published">2004</date><date type="published">2004</date><biblScope unit="vol">43</biblScope><biblScope unit="issue">34</biblScope><biblScope unit="page" from="11103">11103</biblScope><biblScope unit="page" to="11108">11108</biblScope></imprint><idno type="ISSN">0006-2960</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-2960</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome (SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study, the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as shown by 1H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea (with Cm of 2.77 M and m value of 2.74 kcal mol-1 M-1) or GdmCl (with Cm of 1.46 M and m value of 4.50 kcal mol-1 M-1). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to be inactivated. We propose that the low stability of N protein may be critical for the stability and function of SARS virus.</div></front></TEI><istex><corpusName>acs</corpusName><keywords><teeft><json:string>sars</json:string><json:string>nucleocapsid</json:string><json:string>nacl</json:string><json:string>coronavirus</json:string><json:string>sars virus</json:string><json:string>denaturation</json:string><json:string>gdmcl</json:string><json:string>thermostability</json:string><json:string>denatured</json:string><json:string>biochemistry</json:string><json:string>virol</json:string><json:string>viral</json:string><json:string>wang</json:string><json:string>capsid</json:string><json:string>datum</json:string><json:string>anion</json:string><json:string>different concentration</json:string><json:string>final protein concentration</json:string><json:string>nucleocapsid protein</json:string><json:string>urea</json:string><json:string>protein concentration</json:string><json:string>molten globule</json:string><json:string>severe acute respiratory syndrome</json:string><json:string>tris buffer</json:string><json:string>refolding curve</json:string><json:string>protein</json:string><json:string>viral capsid</json:string><json:string>protein denaturation</json:string><json:string>circular dichroism</json:string><json:string>protein increase</json:string><json:string>nacl concentration</json:string><json:string>anion binding</json:string><json:string>important determinant</json:string><json:string>thermal inactivation</json:string><json:string>structural protein</json:string><json:string>stock solution</json:string><json:string>little structure</json:string><json:string>helical nucleocapsid</json:string><json:string>tsinghua university</json:string><json:string>thermal stability</json:string><json:string>acid titration</json:string><json:string>spectroscopic feature</json:string><json:string>maximal emission wavelength</json:string><json:string>emission intensity</json:string><json:string>coronavirus nucleocapsid protein</json:string><json:string>thermostability measurement</json:string><json:string>less stable</json:string><json:string>native state</json:string><json:string>ionic strength</json:string><json:string>gene encoding</json:string><json:string>sodium acetate</json:string><json:string>potassium chloride</json:string><json:string>primary datum</json:string><json:string>microcal software</json:string><json:string>more stable</json:string><json:string>largest increase</json:string><json:string>substantial burial</json:string><json:string>electrostatic interaction</json:string><json:string>native ribonuclease</json:string><json:string>beijing genomics institute</json:string><json:string>major role</json:string><json:string>oral polio vaccine</json:string><json:string>disulfide bond</json:string><json:string>infectious subvirion particle</json:string><json:string>reovirus particle</json:string><json:string>viral rna</json:string><json:string>sars coronavirus</json:string><json:string>inactivation</json:string></teeft></keywords><author><json:item><name>WANG Yulong</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string><json:string>Y.W. and X.W. contributed equally to this study.</json:string></affiliations></json:item><json:item><name>WU Xiaoyu</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string><json:string>Y.W. and X.W. contributed equally to this study.</json:string></affiliations></json:item><json:item><name>WANG Yihua</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string></affiliations></json:item><json:item><name>LI Bing</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string></affiliations></json:item><json:item><name>ZHOU Hao</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string></affiliations></json:item><json:item><name>YUAN Guiyong</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string></affiliations></json:item><json:item><name>FU Yan</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string></affiliations></json:item><json:item><name>LUO Yongzhang</name><affiliations><json:string>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</json:string><json:string>To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.</json:string></affiliations></json:item></author><arkIstex>ark:/67375/TPS-CT10FWKN-F</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome (SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study, the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as shown by 1H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea (with Cm of 2.77 M and m value of 2.74 kcal mol-1 M-1) or GdmCl (with Cm of 1.46 M and m value of 4.50 kcal mol-1 M-1). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to be inactivated. We propose that the low stability of N protein may be critical for the stability and function of SARS virus.</abstract><qualityIndicators><score>9.173</score><pdfWordCount>4449</pdfWordCount><pdfCharCount>25105</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>6</pdfPageCount><pdfPageSize>612 x 792 pts (letter)</pdfPageSize><pdfWordsPerPage>742</pdfWordsPerPage><pdfText>true</pdfText><refBibsNative>false</refBibsNative><abstractWordCount>227</abstractWordCount><abstractCharCount>1346</abstractCharCount><keywordCount>0</keywordCount></qualityIndicators><title>Low Stability of Nucleocapsid Protein in SARS Virus†</title><genre><json:string>research-article</json:string></genre><host><title>Biochemistry</title><language><json:string>unknown</json:string></language><issn><json:string>0006-2960</json:string></issn><eissn><json:string>1520-4995</json:string></eissn><volume>43</volume><issue>34</issue><pages><first>11103</first><last>11108</last></pages><genre><json:string>journal</json:string></genre></host><ark><json:string>ark:/67375/TPS-CT10FWKN-F</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - biochemistry & molecular biology</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - biochemistry & molecular biology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Biochemistry</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences biologiques fondamentales et appliquees. psychologie</json:string></inist></categories><publicationDate>2004</publicationDate><copyrightDate>2004</copyrightDate><doi><json:string>10.1021/bi049194b</json:string></doi><id>096978D90918130B3334B42F12577503A0738C60</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/bundle.zip</uri></json:item><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/fulltext.txt</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main">Low Stability of Nucleocapsid Protein in SARS Virus<ref type="bib" target="#bi049194bAF2">†</ref></title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>American Chemical Society</publisher><availability><licence>Copyright © 2004 American Chemical Society</licence><p>American Chemical Society</p></availability><date type="published">2004</date><date type="Copyright" when="2004">2004</date></publicationStmt><notesStmt><note type="content-type" source="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="publication-type" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note></notesStmt><sourceDesc><biblStruct type="article"><analytic><title level="a" type="main">Low Stability of Nucleocapsid Protein in SARS Virus<ref type="bib" target="#bi049194bAF2">†</ref></title><author xml:id="author-0000"><persName><surname>Wang</surname><forename type="first">Yulong</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation><note place="foot" n="bi049194bAF3"><ref>‡</ref><p>
Y.W. and X.W. contributed equally to this study.</p></note></author><author xml:id="author-0001"><persName><surname>Wu</surname><forename type="first">Xiaoyu</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation><note place="foot" n="bi049194bAF3"><ref>‡</ref><p>
Y.W. and X.W. contributed equally to this study.</p></note></author><author xml:id="author-0002"><persName><surname>Wang</surname><forename type="first">Yihua</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation></author><author xml:id="author-0003"><persName><surname>Li</surname><forename type="first">Bing</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation></author><author xml:id="author-0004"><persName><surname>Zhou</surname><forename type="first">Hao</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation></author><author xml:id="author-0005"><persName><surname>Yuan</surname><forename type="first">Guiyong</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation></author><author xml:id="author-0006"><persName><surname>Fu</surname><forename type="first">Yan</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation></author><author xml:id="author-0007" role="corresp"><persName><surname>Luo</surname><forename type="first">Yongzhang</forename></persName><affiliation><orgName type="laboratory">Department of Biological Sciences and Biotechnology</orgName><orgName type="laboratory">MOE Laboratory of Protein Science</orgName><orgName type="institution">Tsinghua University</orgName><address><addrLine>Beijing 100084</addrLine><addrLine>People's Republic of China</addrLine></address></affiliation><affiliation role="corresp"> To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.</affiliation></author><idno type="istex">096978D90918130B3334B42F12577503A0738C60</idno><idno type="ark">ark:/67375/TPS-CT10FWKN-F</idno><idno type="DOI">10.1021/bi049194b</idno></analytic><monogr><title level="j" type="main">Biochemistry</title><title level="j" type="abbrev">Biochemistry</title><idno type="acspubs">bi</idno><idno type="coden">bichaw</idno><idno type="pISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published">2004</date><date type="published">2004</date><biblScope unit="vol">43</biblScope><biblScope unit="issue">34</biblScope><biblScope unit="page" from="11103">11103</biblScope><biblScope unit="page" to="11108">11108</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><encodingDesc><schemaRef type="ODD" url="https://xml-schema.delivery.istex.fr/tei-istex.odd"></schemaRef><appInfo><application ident="pub2tei" version="1.0.41" when="2020-04-06"><label>pub2TEI-ISTEX</label><desc>A set of style sheets for converting XML documents encoded in various scientific publisher formats into a common TEI format.
<ref target="http://www.tei-c.org/">We use TEI</ref></desc></application></appInfo></encodingDesc><profileDesc><abstract><p><graphic url="bi049194bn00001.tif"></graphic></p><p>The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly
identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome
(SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study,
the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as
shown by <hi rend="superscript">1</hi>H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting
substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little
α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured
near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of
N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea
(with <hi rend="italic">C</hi><hi rend="subscript">m</hi> of 2.77 M and <hi rend="italic">m</hi> value of 2.74 kcal mol<hi rend="superscript">-1</hi> M<hi rend="superscript">-1</hi>) or GdmCl (with <hi rend="italic">C</hi><hi rend="subscript">m</hi> of 1.46 M and <hi rend="italic">m</hi> value of
4.50 kcal mol<hi rend="superscript">-1</hi> M<hi rend="superscript">-1</hi>). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein
starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to
be inactivated. We propose that the low stability of N protein may be critical for the stability and function
of SARS virus.
</p></abstract><textClass ana="subject"><keywords scheme="document-type-name"><term>Article</term></keywords></textClass><langUsage><language ident="en"></language></langUsage></profileDesc><revisionDesc><change when="2020-04-06" who="#istex" xml:id="pub2tei">formatting</change></revisionDesc></teiHeader></istex:fulltextTEI></fulltext><metadata><istex:metadataXml wicri:clean="corpus acs not found" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:document><article article-type="research-article" specific-use="acs2jats-2.0.3"><front><journal-meta><journal-id journal-id-type="acspubs">bi</journal-id><journal-id journal-id-type="coden">bichaw</journal-id><journal-title-group><journal-title>Biochemistry</journal-title><abbrev-journal-title>Biochemistry</abbrev-journal-title></journal-title-group><issn pub-type="ppub">0006-2960</issn><issn pub-type="epub">1520-4995</issn><publisher><publisher-name>American Chemical Society</publisher-name></publisher><self-uri>pubs.acs.org/biochemistry</self-uri></journal-meta><article-meta><article-id pub-id-type="doi">10.1021/bi049194b</article-id><article-categories><subj-group subj-group-type="document-type-name"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Low Stability of Nucleocapsid Protein in SARS Virus<xref rid="bi049194bAF2">†</xref></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Yulong</given-names></name><xref rid="bi049194bAF3">‡</xref></contrib><contrib contrib-type="author"><name><surname>Wu</surname><given-names>Xiaoyu</given-names></name><xref rid="bi049194bAF3">‡</xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Yihua</given-names></name></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Bing</given-names></name></contrib><contrib contrib-type="author"><name><surname>Zhou</surname><given-names>Hao</given-names></name></contrib><contrib contrib-type="author"><name><surname>Yuan</surname><given-names>Guiyong</given-names></name></contrib><contrib contrib-type="author"><name><surname>Fu</surname><given-names>Yan</given-names></name></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Luo</surname><given-names>Yongzhang</given-names></name><xref rid="bi049194bAF1">*</xref></contrib><aff>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,
Beijing 100084, People's Republic of China
</aff></contrib-group><author-notes><fn id="bi049194bAF3"><label>‡</label><p>
Y.W. and X.W. contributed equally to this study.</p></fn><corresp id="bi049194bAF1">
To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.
</corresp></author-notes><pub-date pub-type="epub"><day>6</day><month>08</month><year>2004</year></pub-date><pub-date pub-type="ppub"><day>31</day><month>08</month><year>2004</year></pub-date><volume>43</volume><issue>34</issue><fpage>11103</fpage><lpage>11108</lpage><history><date date-type="received"><day>22</day><month>04</month><year>2004</year></date><date date-type="rev-recd"><day>07</day><month>06</month><year>2004</year></date><date date-type="asap"><day>6</day><month>08</month><year>2004</year></date><date date-type="issue-pub"><day>31</day><month>08</month><year>2004</year></date></history><permissions><copyright-statement>Copyright © 2004 American Chemical Society</copyright-statement><copyright-year>2004</copyright-year><copyright-holder>American Chemical Society</copyright-holder></permissions><abstract><graphic content-type="abstract-graphic" xlink:href="bi049194bn00001.tif"></graphic><p>The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly
identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome
(SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study,
the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as
shown by <sup>1</sup>H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting
substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little
α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured
near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of
N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea
(with <italic>C</italic><sub>m</sub> of 2.77 M and <italic>m</italic> value of 2.74 kcal mol<sup>-1</sup> M<sup>-1</sup>) or GdmCl (with <italic>C</italic><sub>m</sub> of 1.46 M and <italic>m</italic> value of
4.50 kcal mol<sup>-1</sup> M<sup>-1</sup>). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein
starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to
be inactivated. We propose that the low stability of N protein may be critical for the stability and function
of SARS virus.
</p></abstract><custom-meta-group><custom-meta><meta-name>document-id-old-9</meta-name><meta-value>bi049194b</meta-value></custom-meta></custom-meta-group></article-meta><notes id="bi049194bAF2"><label>†</label><p>
Supported in part by grants from Science and Technology Research
Project of the Ministry of Education in China (03007), SARS program
of Beijing Science and Technology Committee (H030230011020), and
SARS Foundation (fd0303) of Tsinghua University.</p></notes></front><body></body><back></back></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Low Stability of Nucleocapsid Protein in SARS Virus†</title></titleInfo><titleInfo contentType="CDATA"><title>Low Stability of Nucleocapsid Protein in SARS Virus†</title></titleInfo><name type="personal"><namePart type="family">WANG</namePart><namePart type="given">Yulong</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><affiliation> Y.W. and X.W. contributed equally to this study.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">WU</namePart><namePart type="given">Xiaoyu</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><affiliation> Y.W. and X.W. contributed equally to this study.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">WANG</namePart><namePart type="given">Yihua</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">LI</namePart><namePart type="given">Bing</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">ZHOU</namePart><namePart type="given">Hao</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">YUAN</namePart><namePart type="given">Guiyong</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">FU</namePart><namePart type="given">Yan</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal" displayLabel="corresp"><namePart type="family">LUO</namePart><namePart type="given">Yongzhang</namePart><affiliation>Department of Biological Sciences and Biotechnology, MOE Laboratory of Protein Science, Tsinghua University,Beijing 100084, People's Republic of China</affiliation><affiliation> To whom correspondence should be addressed. Telephone: 86-10-6277-2897. Fax: 86-10-6279-4691. E-mail: protein@tsinghua.edu.cn.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>American Chemical Society</publisher><dateCreated encoding="w3cdtf">2004-08-06</dateCreated><dateIssued encoding="w3cdtf">2004-08-31</dateIssued><copyrightDate encoding="w3cdtf">2004</copyrightDate></originInfo><note type="footnote" ID="bi049194bAF2"> Supported in part by grants from Science and Technology Research Project of the Ministry of Education in China (03007), SARS program of Beijing Science and Technology Committee (H030230011020), and SARS Foundation (fd0303) of Tsinghua University.</note><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract>The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome (SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study, the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as shown by 1H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little α-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea (with Cm of 2.77 M and m value of 2.74 kcal mol-1 M-1) or GdmCl (with Cm of 1.46 M and m value of 4.50 kcal mol-1 M-1). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein starts to unfold at 35 °C and is completely denatured at 55 °C, where SARS virus was also reported to be inactivated. We propose that the low stability of N protein may be critical for the stability and function of SARS virus.</abstract><note type="footnote" ID="bi049194bAF2"> Supported in part by grants from Science and Technology Research Project of the Ministry of Education in China (03007), SARS program of Beijing Science and Technology Committee (H030230011020), and SARS Foundation (fd0303) of Tsinghua University.</note><relatedItem type="host"><titleInfo><title>Biochemistry</title></titleInfo><titleInfo type="abbreviated"><title>Biochemistry</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><identifier type="ISSN">0006-2960</identifier><identifier type="eISSN">1520-4995</identifier><identifier type="acspubs">bi</identifier><identifier type="coden">BICHAW</identifier><identifier type="uri">pubs.acs.org/biochemistry</identifier><part><date>2004</date><detail type="volume"><caption>vol.</caption><number>43</number></detail><detail type="issue"><caption>no.</caption><number>34</number></detail><extent unit="pages"><start>11103</start><end>11108</end></extent></part></relatedItem><identifier type="istex">096978D90918130B3334B42F12577503A0738C60</identifier><identifier type="ark">ark:/67375/TPS-CT10FWKN-F</identifier><identifier type="DOI">10.1021/bi049194b</identifier><accessCondition type="use and reproduction" contentType="restricted">Copyright © 2004 American Chemical Society</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-X5HBJWF8-J">acs</recordContentSource><recordOrigin>Converted from (version 1.2.10) to MODS version 3.6.</recordOrigin><recordCreationDate encoding="w3cdtf">2020-04-10</recordCreationDate></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-CT10FWKN-F/record.json</uri></json:item></metadata><annexes><json:item><extension>tiff</extension><original>true</original><mimetype>image/tiff</mimetype><uri>https://api.istex.fr/document/096978D90918130B3334B42F12577503A0738C60/annexes/tiff</uri></json:item></annexes><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/StressCovidV1/Data/Istex/Corpus
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000009 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Corpus/biblio.hfd -nk 000009 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= StressCovidV1
|flux= Istex
|étape= Corpus
|type= RBID
|clé= ISTEX:096978D90918130B3334B42F12577503A0738C60
|texte= Low Stability of Nucleocapsid Protein in SARS Virus†
}}
| This area was generated with Dilib version V0.6.33. |  |