Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride
Identifieur interne : 000F08 ( Pmc/Checkpoint ); précédent : 000F07; suivant : 000F09Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride
Auteurs : Hui-Ping Chang ; Chi-Yuan Chou ; Gu-Gang ChangSource :
- Biophysical Journal [ 0006-3495 ] ; 2007.
Abstract
Chemical denaturant sensitivity of the dimeric main protease from severe acute respiratory syndrome (SARS) coronavirus to guanidinium chloride was examined in terms of fluorescence spectroscopy, circular dichroism, analytical ultracentrifuge, and enzyme activity change. The dimeric enzyme dissociated at guanidinium chloride concentration of <0.4 M, at which the enzymatic activity loss showed close correlation with the subunit dissociation. Further increase in guanidinium chloride induced a reversible biphasic unfolding of the enzyme. The unfolding of the C-terminal domain-truncated enzyme, on the other hand, followed a monophasic unfolding curve. Different mutants of the full-length protease (W31 and W207/W218), with tryptophanyl residue(s) mutated to phenylalanine at the C-terminal or N-terminal domain, respectively, were constructed. Unfolding curves of these mutants were monophasic but corresponded to the first and second phases of the protease, respectively. The unfolding intermediate of the protease thus represented a folded C-terminal domain but an unfolded N-terminal domain, which is enzymatically inactive due to loss of regulatory properties. The various enzyme forms were characterized in terms of hydrophobicity and size-and-shape distributions. We provide direct evidence for the functional role of C-terminal domain in stabilization of the catalytic N-terminal domain of SARS coronavirus main protease.
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DOI: 10.1529/biophysj.106.091736
PubMed: 17142288
PubMed Central: 1783898
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride</title><author><name sortKey="Chang, Hui Ping" sort="Chang, Hui Ping" uniqKey="Chang H" first="Hui-Ping" last="Chang">Hui-Ping Chang</name></author><author><name sortKey="Chou, Chi Yuan" sort="Chou, Chi Yuan" uniqKey="Chou C" first="Chi-Yuan" last="Chou">Chi-Yuan Chou</name></author><author><name sortKey="Chang, Gu Gang" sort="Chang, Gu Gang" uniqKey="Chang G" first="Gu-Gang" last="Chang">Gu-Gang Chang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">17142288</idno><idno type="pmc">1783898</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1783898</idno><idno type="RBID">PMC:1783898</idno><idno type="doi">10.1529/biophysj.106.091736</idno><date when="2007">2007</date><idno type="wicri:Area/Pmc/Corpus">000E55</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000E55</idno><idno type="wicri:Area/Pmc/Curation">000E55</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000E55</idno><idno type="wicri:Area/Pmc/Checkpoint">000F08</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000F08</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride</title><author><name sortKey="Chang, Hui Ping" sort="Chang, Hui Ping" uniqKey="Chang H" first="Hui-Ping" last="Chang">Hui-Ping Chang</name></author><author><name sortKey="Chou, Chi Yuan" sort="Chou, Chi Yuan" uniqKey="Chou C" first="Chi-Yuan" last="Chou">Chi-Yuan Chou</name></author><author><name sortKey="Chang, Gu Gang" sort="Chang, Gu Gang" uniqKey="Chang G" first="Gu-Gang" last="Chang">Gu-Gang Chang</name></author></analytic><series><title level="j">Biophysical Journal</title><idno type="ISSN">0006-3495</idno><idno type="eISSN">1542-0086</idno><imprint><date when="2007">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Chemical denaturant sensitivity of the dimeric main protease from severe acute respiratory syndrome (SARS) coronavirus to guanidinium chloride was examined in terms of fluorescence spectroscopy, circular dichroism, analytical ultracentrifuge, and enzyme activity change. The dimeric enzyme dissociated at guanidinium chloride concentration of <0.4 M, at which the enzymatic activity loss showed close correlation with the subunit dissociation. Further increase in guanidinium chloride induced a reversible biphasic unfolding of the enzyme. The unfolding of the C-terminal domain-truncated enzyme, on the other hand, followed a monophasic unfolding curve. Different mutants of the full-length protease (W31 and W207/W218), with tryptophanyl residue(s) mutated to phenylalanine at the C-terminal or N-terminal domain, respectively, were constructed. Unfolding curves of these mutants were monophasic but corresponded to the first and second phases of the protease, respectively. The unfolding intermediate of the protease thus represented a folded C-terminal domain but an unfolded N-terminal domain, which is enzymatically inactive due to loss of regulatory properties. The various enzyme forms were characterized in terms of hydrophobicity and size-and-shape distributions. We provide direct evidence for the functional role of C-terminal domain in stabilization of the catalytic N-terminal domain of SARS coronavirus main protease.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Chan, H L" uniqKey="Chan H">H.L. Chan</name></author><author><name sortKey="Tsui, S K" uniqKey="Tsui S">S.K. Tsui</name></author><author><name sortKey="Sung, J J" uniqKey="Sung J">J.J. Sung</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Matsuyama, S" uniqKey="Matsuyama S">S. Matsuyama</name></author><author><name sortKey="Ujike, M" uniqKey="Ujike M">M. Ujike</name></author><author><name sortKey="Morikawa, S" uniqKey="Morikawa S">S. Morikawa</name></author><author><name sortKey="Tashiro, M" uniqKey="Tashiro M">M. Tashiro</name></author><author><name sortKey="Taguchi, F" uniqKey="Taguchi F">F. Taguchi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yang, H" uniqKey="Yang H">H. Yang</name></author><author><name sortKey="Xie, W" uniqKey="Xie W">W. Xie</name></author><author><name sortKey="Xue, X" uniqKey="Xue X">X. Xue</name></author><author><name sortKey="Yang, K" uniqKey="Yang K">K. Yang</name></author><author><name sortKey="Ma, J" uniqKey="Ma J">J. Ma</name></author><author><name sortKey="Liang, W" uniqKey="Liang W">W. Liang</name></author><author><name sortKey="Zhao, Q" uniqKey="Zhao Q">Q. Zhao</name></author><author><name sortKey="Zhou, Z" uniqKey="Zhou Z">Z. Zhou</name></author><author><name sortKey="Pei, D" uniqKey="Pei D">D. Pei</name></author><author><name sortKey="Ziebuhr, J" uniqKey="Ziebuhr J">J. Ziebuhr</name></author><author><name sortKey="Hilgenfeld, R" uniqKey="Hilgenfeld R">R. Hilgenfeld</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K.Y. Yuen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wei, P" uniqKey="Wei P">P. Wei</name></author><author><name sortKey="Fan, K" uniqKey="Fan K">K. Fan</name></author><author><name sortKey="Chen, H" uniqKey="Chen H">H. Chen</name></author><author><name sortKey="Ma, L" uniqKey="Ma L">L. Ma</name></author><author><name sortKey="Huang, C" uniqKey="Huang C">C. Huang</name></author><author><name sortKey="Tan, L" uniqKey="Tan L">L. Tan</name></author><author><name sortKey="Xi, D" uniqKey="Xi D">D. Xi</name></author><author><name sortKey="Li, C" uniqKey="Li C">C. Li</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Cao, A" uniqKey="Cao A">A. Cao</name></author><author><name sortKey="Lai, L" uniqKey="Lai L">L. Lai</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liang, P H" uniqKey="Liang P">P.H. Liang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tong, L" uniqKey="Tong L">L. Tong</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yang, H" uniqKey="Yang H">H. Yang</name></author><author><name sortKey="Yang, M" uniqKey="Yang M">M. Yang</name></author><author><name sortKey="Ding, Y" uniqKey="Ding Y">Y. Ding</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Lou, Z" uniqKey="Lou Z">Z. Lou</name></author><author><name sortKey="Zhou, Z" uniqKey="Zhou Z">Z. Zhou</name></author><author><name sortKey="Sun, L" uniqKey="Sun L">L. Sun</name></author><author><name sortKey="Mo, L" uniqKey="Mo L">L. Mo</name></author><author><name sortKey="Ye, S" uniqKey="Ye S">S. Ye</name></author><author><name sortKey="Pang, H" uniqKey="Pang H">H. Pang</name></author><author><name sortKey="Gao, G F" uniqKey="Gao G">G.F. Gao</name></author><author><name sortKey="Anand, K" uniqKey="Anand K">K. Anand</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bartlam, M" uniqKey="Bartlam M">M. Bartlam</name></author><author><name sortKey="Yang, H" uniqKey="Yang H">H. Yang</name></author><author><name sortKey="Rao, Z" uniqKey="Rao Z">Z. Rao</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Anand, K" uniqKey="Anand K">K. Anand</name></author><author><name sortKey="Palm, G J" uniqKey="Palm G">G.J. Palm</name></author><author><name sortKey="Mesters, J R" uniqKey="Mesters J">J.R. Mesters</name></author><author><name sortKey="Siddell, S G" uniqKey="Siddell S">S.G. Siddell</name></author><author><name sortKey="Ziebuhr, J" uniqKey="Ziebuhr J">J. Ziebuhr</name></author><author><name sortKey="Hilgenfeld, R" uniqKey="Hilgenfeld R">R. Hilgenfeld</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Anand, K" uniqKey="Anand K">K. Anand</name></author><author><name sortKey="Ziebuhr, J" uniqKey="Ziebuhr J">J. Ziebuhr</name></author><author><name sortKey="Wadhwani, P" uniqKey="Wadhwani P">P. Wadhwani</name></author><author><name sortKey="Mesters, J R" uniqKey="Mesters J">J.R. Mesters</name></author><author><name sortKey="Hilgenfeld, R" uniqKey="Hilgenfeld R">R. Hilgenfeld</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chou, C Y" uniqKey="Chou C">C.Y. Chou</name></author><author><name sortKey="Chang, H C" uniqKey="Chang H">H.C. Chang</name></author><author><name sortKey="Hsu, W C" uniqKey="Hsu W">W.C. Hsu</name></author><author><name sortKey="Lin, T Z" uniqKey="Lin T">T.Z. Lin</name></author><author><name sortKey="Lin, C H" uniqKey="Lin C">C.H. Lin</name></author><author><name sortKey="Chang, G G" uniqKey="Chang G">G.G. Chang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hsu, W C" uniqKey="Hsu W">W.C. Hsu</name></author><author><name sortKey="Chang, H C" uniqKey="Chang H">H.C. Chang</name></author><author><name sortKey="Chou, C Y" uniqKey="Chou C">C.Y. Chou</name></author><author><name sortKey="Tsai, P J" uniqKey="Tsai P">P.J. Tsai</name></author><author><name sortKey="Lin, P I" uniqKey="Lin P">P.I. Lin</name></author><author><name sortKey="Chang, G G" uniqKey="Chang G">G.G. Chang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hsu, M F" uniqKey="Hsu M">M.F. Hsu</name></author><author><name sortKey="Kuo, C J" uniqKey="Kuo C">C.J. Kuo</name></author><author><name sortKey="Chang, K T" uniqKey="Chang K">K.T. Chang</name></author><author><name sortKey="Chang, H C" uniqKey="Chang H">H.C. Chang</name></author><author><name sortKey="Chou, C C" uniqKey="Chou C">C.C. Chou</name></author><author><name sortKey="Ko, T P" uniqKey="Ko T">T.P. Ko</name></author><author><name sortKey="Shr, H L" uniqKey="Shr H">H.L. Shr</name></author><author><name sortKey="Chang, G G" uniqKey="Chang G">G.G. Chang</name></author><author><name sortKey="Wang, A H" uniqKey="Wang A">A.H. Wang</name></author><author><name sortKey="Liang, P H" uniqKey="Liang P">P.H. Liang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Braman, J" uniqKey="Braman J">J. Braman</name></author><author><name sortKey="Papworth, C" uniqKey="Papworth C">C. Papworth</name></author><author><name sortKey="Greener, A" uniqKey="Greener A">A. Greener</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Royer, C A" uniqKey="Royer C">C.A. Royer</name></author><author><name sortKey="Mann, C J" uniqKey="Mann C">C.J. Mann</name></author><author><name sortKey="Matthews, C R" uniqKey="Matthews C">C.R. Matthews</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Schuck, P" uniqKey="Schuck P">P. Schuck</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Brown, P H" uniqKey="Brown P">P.H. Brown</name></author><author><name sortKey="Schuck, P" uniqKey="Schuck P">P. Schuck</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lamm, O" uniqKey="Lamm O">O. Lamm</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pace, C N" uniqKey="Pace C">C.N. Pace</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Morjana, N A" uniqKey="Morjana N">N.A. Morjana</name></author><author><name sortKey="Mckeone, B J" uniqKey="Mckeone B">B.J. McKeone</name></author><author><name sortKey="Gilbert, H F" uniqKey="Gilbert H">H.F. Gilbert</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chen, H" uniqKey="Chen H">H. Chen</name></author><author><name sortKey="Wei, P" uniqKey="Wei P">P. Wei</name></author><author><name sortKey="Huang, C" uniqKey="Huang C">C. Huang</name></author><author><name sortKey="Tan, L" uniqKey="Tan L">L. Tan</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Lai, L" uniqKey="Lai L">L. Lai</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Laskowski, R A" uniqKey="Laskowski R">R.A. Laskowski</name></author><author><name sortKey="Macarthur, M W" uniqKey="Macarthur M">M.W. MacArthur</name></author><author><name sortKey="Moss, D S" uniqKey="Moss D">D.S. Moss</name></author><author><name sortKey="Thornton, J M" uniqKey="Thornton J">J.M. Thornton</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, T W" uniqKey="Lee T">T.W. Lee</name></author><author><name sortKey="Cherney, M M" uniqKey="Cherney M">M.M. Cherney</name></author><author><name sortKey="Huitema, C" uniqKey="Huitema C">C. Huitema</name></author><author><name sortKey="Liu, J" uniqKey="Liu J">J. Liu</name></author><author><name sortKey="James, K E" uniqKey="James K">K.E. James</name></author><author><name sortKey="Powers, J C" uniqKey="Powers J">J.C. Powers</name></author><author><name sortKey="Eltis, L D" uniqKey="Eltis L">L.D. Eltis</name></author><author><name sortKey="James, M N" uniqKey="James M">M.N. James</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bacha, U" uniqKey="Bacha U">U. Bacha</name></author><author><name sortKey="Barrila, J" uniqKey="Barrila J">J. Barrila</name></author><author><name sortKey="Velazquez Campoy, A" uniqKey="Velazquez Campoy A">A. Velazquez-Campoy</name></author><author><name sortKey="Leavitt, S A" uniqKey="Leavitt S">S.A. Leavitt</name></author><author><name sortKey="Freire, E" uniqKey="Freire E">E. Freire</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shi, J" uniqKey="Shi J">J. Shi</name></author><author><name sortKey="Wei, Z" uniqKey="Wei Z">Z. Wei</name></author><author><name sortKey="Song, J" uniqKey="Song J">J. Song</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shortle, D" uniqKey="Shortle D">D. Shortle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fink, A L" uniqKey="Fink A">A.L. Fink</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hegyi, A" uniqKey="Hegyi A">A. Hegyi</name></author><author><name sortKey="Friebe, A" uniqKey="Friebe A">A. Friebe</name></author><author><name sortKey="Gorbalenya, A E" uniqKey="Gorbalenya A">A.E. Gorbalenya</name></author><author><name sortKey="Ziebuhr, J" uniqKey="Ziebuhr J">J. Ziebuhr</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Huang, C" uniqKey="Huang C">C. Huang</name></author><author><name sortKey="Wei, P" uniqKey="Wei P">P. Wei</name></author><author><name sortKey="Fan, K" uniqKey="Fan K">K. Fan</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Lai, L" uniqKey="Lai L">L. Lai</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tan, J" uniqKey="Tan J">J. Tan</name></author><author><name sortKey="Verschueren, K H" uniqKey="Verschueren K">K.H. Verschueren</name></author><author><name sortKey="Anand, K" uniqKey="Anand K">K. Anand</name></author><author><name sortKey="Shen, J" uniqKey="Shen J">J. Shen</name></author><author><name sortKey="Yang, M" uniqKey="Yang M">M. Yang</name></author><author><name sortKey="Xu, Y" uniqKey="Xu Y">Y. Xu</name></author><author><name sortKey="Rao, Z" uniqKey="Rao Z">Z. Rao</name></author><author><name sortKey="Bigalke, J" uniqKey="Bigalke J">J. Bigalke</name></author><author><name sortKey="Heisen, B" uniqKey="Heisen B">B. Heisen</name></author><author><name sortKey="Mesters, J R" uniqKey="Mesters J">J.R. Mesters</name></author><author><name sortKey="Chen, K" uniqKey="Chen K">K. Chen</name></author><author><name sortKey="Shen, X" uniqKey="Shen X">X. Shen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dill, K A" uniqKey="Dill K">K.A. Dill</name></author><author><name sortKey="Chan, H S" uniqKey="Chan H">H.S. Chan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jahn, T R" uniqKey="Jahn T">T.R. Jahn</name></author><author><name sortKey="Radford, S E" uniqKey="Radford S">S.E. Radford</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gruebele, M" uniqKey="Gruebele M">M. Gruebele</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lindorff Larsen, K" uniqKey="Lindorff Larsen K">K. Lindorff-Larsen</name></author><author><name sortKey="Rogen, P" uniqKey="Rogen P">P. Rogen</name></author><author><name sortKey="Paci, E" uniqKey="Paci E">E. Paci</name></author><author><name sortKey="Vendruscolo, M" uniqKey="Vendruscolo M">M. Vendruscolo</name></author><author><name sortKey="Dobson, C M" uniqKey="Dobson C">C.M. Dobson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Oliveberg, M" uniqKey="Oliveberg M">M. Oliveberg</name></author><author><name sortKey="Shakhnovich, E I" uniqKey="Shakhnovich E">E.I. Shakhnovich</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shi, J" uniqKey="Shi J">J. Shi</name></author><author><name sortKey="Song, J" uniqKey="Song J">J. Song</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Delano, W L" uniqKey="Delano W">W.L. DeLano</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Biophys J</journal-id><journal-id journal-id-type="iso-abbrev">Biophys. J</journal-id><journal-title-group><journal-title>Biophysical Journal</journal-title></journal-title-group><issn pub-type="ppub">0006-3495</issn><issn pub-type="epub">1542-0086</issn><publisher><publisher-name>The Biophysical Society. Published by Elsevier Inc.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">17142288</article-id><article-id pub-id-type="pmc">1783898</article-id><article-id pub-id-type="publisher-id">S0006-3495(07)70948-7</article-id><article-id pub-id-type="doi">10.1529/biophysj.106.091736</article-id><article-categories><subj-group subj-group-type="heading"><subject>Proteins</subject></subj-group></article-categories><title-group><article-title>Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Chang</surname><given-names>Hui-Ping</given-names></name><email>huiping_chp@hotmail.com</email><xref rid="cor1" ref-type="corresp">⁎</xref></contrib><contrib contrib-type="author"><name><surname>Chou</surname><given-names>Chi-Yuan</given-names></name></contrib><contrib contrib-type="author"><name><surname>Chang</surname><given-names>Gu-Gang</given-names></name></contrib></contrib-group><aff>Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan</aff><author-notes><corresp id="cor1"><label>⁎</label>Address reprint requests to Hui-Ping Chang, Dept. of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, 155 Li-Nong St., Section 2, Taipei 112, Taiwan. <email>huiping_chp@hotmail.com</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>6</day><month>1</month><year>2009</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><day>15</day><month>2</month><year>2007</year></pub-date><pub-date pub-type="epub"><day>6</day><month>1</month><year>2009</year></pub-date><volume>92</volume><issue>4</issue><fpage>1374</fpage><lpage>1383</lpage><history><date date-type="received"><day>19</day><month>6</month><year>2006</year></date><date date-type="accepted"><day>2</day><month>11</month><year>2006</year></date></history><permissions><copyright-statement>Copyright © 2007 The Biophysical Society. Published by Elsevier Inc. All rights reserved.</copyright-statement><copyright-year>2007</copyright-year><copyright-holder>The Biophysical Society</copyright-holder><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract><p>Chemical denaturant sensitivity of the dimeric main protease from severe acute respiratory syndrome (SARS) coronavirus to guanidinium chloride was examined in terms of fluorescence spectroscopy, circular dichroism, analytical ultracentrifuge, and enzyme activity change. The dimeric enzyme dissociated at guanidinium chloride concentration of <0.4 M, at which the enzymatic activity loss showed close correlation with the subunit dissociation. Further increase in guanidinium chloride induced a reversible biphasic unfolding of the enzyme. The unfolding of the C-terminal domain-truncated enzyme, on the other hand, followed a monophasic unfolding curve. Different mutants of the full-length protease (W31 and W207/W218), with tryptophanyl residue(s) mutated to phenylalanine at the C-terminal or N-terminal domain, respectively, were constructed. Unfolding curves of these mutants were monophasic but corresponded to the first and second phases of the protease, respectively. The unfolding intermediate of the protease thus represented a folded C-terminal domain but an unfolded N-terminal domain, which is enzymatically inactive due to loss of regulatory properties. The various enzyme forms were characterized in terms of hydrophobicity and size-and-shape distributions. We provide direct evidence for the functional role of C-terminal domain in stabilization of the catalytic N-terminal domain of SARS coronavirus main protease.</p></abstract><kwd-group><title>Abbreviations used</title><kwd>SARS, severe acute respiratory syndrome</kwd><kwd>CoV, coronavirus</kwd><kwd>3CLpro, 3-chymotrypsin-like SARS-CoV main protease</kwd><kwd>3CL<sub>I+II</sub>, C-terminal domain-truncated SARS-CoV main protease</kwd><kwd>W31 (or more precisely W207F/W218F), double mutant of SARS-CoV 3CLpro with tryptophanyl residues at 207 and 218 mutated to phenylalanine</kwd><kwd>W207/W218 (or more precisely W31F), point mutant of SARS-CoV 3CLpro with tryptophanyl residue at position 31 mutated to phenylalanine</kwd><kwd>AEW, average emission wavelength</kwd><kwd>CD, circular dichroism</kwd><kwd>AUC, analytical ultracentrifugation</kwd><kwd>GdnCl, guanidinium chloride</kwd><kwd><italic>f/f</italic><sub>o</sub>, frictional ratio</kwd><kwd>ANS, 1-anilino-8-naphthalene sulfonic acid</kwd></kwd-group></article-meta></front></pmc><affiliations><list></list><tree><noCountry><name sortKey="Chang, Gu Gang" sort="Chang, Gu Gang" uniqKey="Chang G" first="Gu-Gang" last="Chang">Gu-Gang Chang</name><name sortKey="Chang, Hui Ping" sort="Chang, Hui Ping" uniqKey="Chang H" first="Hui-Ping" last="Chang">Hui-Ping Chang</name><name sortKey="Chou, Chi Yuan" sort="Chou, Chi Yuan" uniqKey="Chou C" first="Chi-Yuan" last="Chou">Chi-Yuan Chou</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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