Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†
Identifieur interne : 000045 ( Istex/Corpus ); précédent : 000044; suivant : 000046Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†
Auteurs : Changying Yu ; Chunshan Gui ; Haibin Luo ; Lili Chen ; Liang Zhang ; Hao Yu ; Sheng Yang ; Weihong Jiang ; Jianhua Shen ; Xu Shen ; Hualiang JiangSource :
- Biochemistry [ 0006-2960 ] ; 2005.
Abstract
Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus (SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques. For the thermally induced unfolding at pH 7.4, the apparent activation energy (Eapp) and transition midpoint temperature (Tm) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent spectra resulted in consistent values of the transition middle concentration of the denaturant (Cm, ranging from 2.30 to 2.45 M) and the standard free energy change (ΔG°, ranging from 2.1 to 2.5 kcal/mol) for the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding reaction rates and relaxation times were determined at various GuHCl concentrations, and the Cm value was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations. Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional model was also used to explain the recently published experimental results of S1−ACE-2 binding and immunizations.
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DOI: 10.1021/bi0482396
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†</title><author><name sortKey="Yu, Changying" sort="Yu, Changying" uniqKey="Yu C" first="Changying" last="Yu">Changying Yu</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> These authors contributed equally to this work.</mods:affiliation></affiliation></author><author><name sortKey="Gui, Chunshan" sort="Gui, Chunshan" uniqKey="Gui C" first="Chunshan" last="Gui">Chunshan Gui</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> These authors contributed equally to this work.</mods:affiliation></affiliation></author><author><name sortKey="Luo, Haibin" sort="Luo, Haibin" uniqKey="Luo H" first="Haibin" last="Luo">Haibin Luo</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Chen, Lili" sort="Chen, Lili" uniqKey="Chen L" first="Lili" last="Chen">Lili Chen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Zhang, Liang" sort="Zhang, Liang" uniqKey="Zhang L" first="Liang" last="Zhang">Liang Zhang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Yu, Hao" sort="Yu, Hao" uniqKey="Yu H" first="Hao" last="Yu">Hao Yu</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Yang, Sheng" sort="Yang, Sheng" uniqKey="Yang S" first="Sheng" last="Yang">Sheng Yang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Jiang, Weihong" sort="Jiang, Weihong" uniqKey="Jiang W" first="Weihong" last="Jiang">Weihong Jiang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Shen, Jianhua" sort="Shen, Jianhua" uniqKey="Shen J" first="Jianhua" last="Shen">Jianhua Shen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Shen, Xu" sort="Shen, Xu" uniqKey="Shen X" first="Xu" last="Shen">Xu Shen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</mods:affiliation></affiliation></author><author><name sortKey="Jiang, Hualiang" sort="Jiang, Hualiang" uniqKey="Jiang H" first="Hualiang" last="Jiang">Hualiang Jiang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> East-China University of Science and Technology.</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052</idno><date when="2005" year="2005">2005</date><idno type="doi">10.1021/bi0482396</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-NZ5V5XNP-C/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000045</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000045</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment
(SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with
Three-Dimensional Structural Modeling<ref type="bib" target="#bi0482396AF2">†</ref></title><author><name sortKey="Yu, Changying" sort="Yu, Changying" uniqKey="Yu C" first="Changying" last="Yu">Changying Yu</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> These authors contributed equally to this work.</mods:affiliation></affiliation></author><author><name sortKey="Gui, Chunshan" sort="Gui, Chunshan" uniqKey="Gui C" first="Chunshan" last="Gui">Chunshan Gui</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> These authors contributed equally to this work.</mods:affiliation></affiliation></author><author><name sortKey="Luo, Haibin" sort="Luo, Haibin" uniqKey="Luo H" first="Haibin" last="Luo">Haibin Luo</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Chen, Lili" sort="Chen, Lili" uniqKey="Chen L" first="Lili" last="Chen">Lili Chen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Zhang, Liang" sort="Zhang, Liang" uniqKey="Zhang L" first="Liang" last="Zhang">Liang Zhang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Yu, Hao" sort="Yu, Hao" uniqKey="Yu H" first="Hao" last="Yu">Hao Yu</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Yang, Sheng" sort="Yang, Sheng" uniqKey="Yang S" first="Sheng" last="Yang">Sheng Yang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Jiang, Weihong" sort="Jiang, Weihong" uniqKey="Jiang W" first="Weihong" last="Jiang">Weihong Jiang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Shen, Jianhua" sort="Shen, Jianhua" uniqKey="Shen J" first="Jianhua" last="Shen">Jianhua Shen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation></author><author><name sortKey="Shen, Xu" sort="Shen, Xu" uniqKey="Shen X" first="Xu" last="Shen">Xu Shen</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</mods:affiliation></affiliation></author><author><name sortKey="Jiang, Hualiang" sort="Jiang, Hualiang" uniqKey="Jiang H" first="Hualiang" last="Jiang">Hualiang Jiang</name><affiliation><mods:affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</mods:affiliation></affiliation><affiliation><mods:affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</mods:affiliation></affiliation><affiliation><mods:affiliation> East-China University of Science and Technology.</mods:affiliation></affiliation><affiliation><mods:affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</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">2005</date><date type="published">2005</date><biblScope unit="vol">44</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="1453">1453</biblScope><biblScope unit="page" to="1463">1463</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">Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus (SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques. For the thermally induced unfolding at pH 7.4, the apparent activation energy (Eapp) and transition midpoint temperature (Tm) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent spectra resulted in consistent values of the transition middle concentration of the denaturant (Cm, ranging from 2.30 to 2.45 M) and the standard free energy change (ΔG°, ranging from 2.1 to 2.5 kcal/mol) for the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding reaction rates and relaxation times were determined at various GuHCl concentrations, and the Cm value was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations. Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional model was also used to explain the recently published experimental results of S1−ACE-2 binding and immunizations.</div></front></TEI><istex><corpusName>acs</corpusName><keywords><teeft><json:string>coronavirus</json:string><json:string>sars</json:string><json:string>denaturant</json:string><json:string>refolding</json:string><json:string>denaturation</json:string><json:string>guhcl</json:string><json:string>tryptophan</json:string><json:string>shanghai</json:string><json:string>biochemistry</json:string><json:string>shen</json:string><json:string>datum</json:string><json:string>relaxation time</json:string><json:string>severe acute respiratory syndrome</json:string><json:string>glycoprotein</json:string><json:string>shanghai institute</json:string><json:string>jiang</json:string><json:string>dichroism</json:string><json:string>immunodominant</json:string><json:string>wong</json:string><json:string>subunit</json:string><json:string>kinetic 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decrease</json:string><json:string>equilibrium condition</json:string><json:string>emission fluorescence</json:string><json:string>refolding process</json:string><json:string>shanghai science</json:string><json:string>thermodynamic parameter</json:string><json:string>different volume ratio</json:string><json:string>refolding procedure</json:string><json:string>quartz cell</json:string><json:string>technology commission</json:string><json:string>sigmoidal method</json:string><json:string>fluorescence emission</json:string><json:string>neslab water bath</json:string><json:string>datum analysis</json:string><json:string>steady state</json:string><json:string>immunological fragment</json:string><json:string>binding receptor</json:string><json:string>sars diagnostics</json:string><json:string>acid fragment</json:string><json:string>mutation site</json:string><json:string>side chain</json:string><json:string>weblab viewerpro</json:string><json:string>hong kong</json:string><json:string>scan rate</json:string><json:string>acta pharmacol</json:string><json:string>mass spectrometric characterization</json:string><json:string>jasco spectropolarimeter</json:string></teeft></keywords><author><json:item><name>YU Changying</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string><json:string>These authors contributed equally to this work.</json:string></affiliations></json:item><json:item><name>GUI Chunshan</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string><json:string>These authors contributed equally to this work.</json:string></affiliations></json:item><json:item><name>LUO Haibin</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string></affiliations></json:item><json:item><name>CHEN Lili</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string></affiliations></json:item><json:item><name>ZHANG Liang</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string></affiliations></json:item><json:item><name>YU Hao</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</json:string></affiliations></json:item><json:item><name>YANG Sheng</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</json:string></affiliations></json:item><json:item><name>JIANG Weihong</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</json:string></affiliations></json:item><json:item><name>SHEN Jianhua</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string></affiliations></json:item><json:item><name>SHEN Xu</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string><json:string>To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</json:string></affiliations></json:item><json:item><name>JIANG Hualiang</name><affiliations><json:string>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</json:string><json:string>Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</json:string><json:string>East-China University of Science and Technology.</json:string><json:string>To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</json:string></affiliations></json:item></author><arkIstex>ark:/67375/TPS-NZ5V5XNP-C</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus (SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques. For the thermally induced unfolding at pH 7.4, the apparent activation energy (Eapp) and transition midpoint temperature (Tm) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent spectra resulted in consistent values of the transition middle concentration of the denaturant (Cm, ranging from 2.30 to 2.45 M) and the standard free energy change (ΔG°, ranging from 2.1 to 2.5 kcal/mol) for the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding reaction rates and relaxation times were determined at various GuHCl concentrations, and the Cm value was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations. Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional model was also used to explain the recently published experimental results of S1−ACE-2 binding and immunizations.</abstract><qualityIndicators><score>10</score><pdfWordCount>7393</pdfWordCount><pdfCharCount>43686</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>11</pdfPageCount><pdfPageSize>612 x 792 pts (letter)</pdfPageSize><pdfWordsPerPage>672</pdfWordsPerPage><pdfText>true</pdfText><refBibsNative>false</refBibsNative><abstractWordCount>278</abstractWordCount><abstractCharCount>1949</abstractCharCount><keywordCount>0</keywordCount></qualityIndicators><title>Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†</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>44</volume><issue>5</issue><pages><first>1453</first><last>1463</last></pages><genre><json:string>journal</json:string></genre></host><ark><json:string>ark:/67375/TPS-NZ5V5XNP-C</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>2005</publicationDate><copyrightDate>2005</copyrightDate><doi><json:string>10.1021/bi0482396</json:string></doi><id>1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052</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-NZ5V5XNP-C/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-NZ5V5XNP-C/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-NZ5V5XNP-C/fulltext.txt</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/TPS-NZ5V5XNP-C/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main">Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment
(SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with
Three-Dimensional Structural Modeling<ref type="bib" target="#bi0482396AF2">†</ref></title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>American Chemical Society</publisher><availability><licence>Copyright © 2005 American Chemical Society</licence><p>American Chemical Society</p></availability><date type="published">2005</date><date type="Copyright" when="2005">2005</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">Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment
(SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with
Three-Dimensional Structural Modeling<ref type="bib" target="#bi0482396AF2">†</ref></title><author xml:id="author-0000"><persName><surname>Yu</surname><forename type="first">Changying</forename></persName><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note><note place="foot" n="bi0482396AF4"><ref>§</ref><p>
These authors contributed equally to this work.</p></note></author><author xml:id="author-0001"><persName><surname>Gui</surname><forename type="first">Chunshan</forename></persName><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note><note place="foot" n="bi0482396AF4"><ref>§</ref><p>
These authors contributed equally to this work.</p></note></author><author xml:id="author-0002"><persName><surname>Luo</surname><forename type="first">Haibin</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note></author><author xml:id="author-0003"><persName><surname>Chen</surname><forename type="first">Lili</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note></author><author xml:id="author-0004"><persName><surname>Zhang</surname><forename type="first">Liang</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note></author><author xml:id="author-0005"><persName><surname>Yu</surname><forename type="first">Hao</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF5"><ref>‖</ref><p>
Shanghai Institute of Plant Physiology & Ecology, Shanghai
Institutes for Biological Sciences, Graduate School, Chinese Academy
of Sciences.</p></note></author><author xml:id="author-0006"><persName><surname>Yang</surname><forename type="first">Sheng</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF5"><ref>‖</ref><p>
Shanghai Institute of Plant Physiology & Ecology, Shanghai
Institutes for Biological Sciences, Graduate School, Chinese Academy
of Sciences.</p></note></author><author xml:id="author-0007"><persName><surname>Jiang</surname><forename type="first">Weihong</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF5"><ref>‖</ref><p>
Shanghai Institute of Plant Physiology & Ecology, Shanghai
Institutes for Biological Sciences, Graduate School, Chinese Academy
of Sciences.</p></note></author><author xml:id="author-0008"><persName><surname>Shen</surname><forename type="first">Jianhua</forename></persName><affiliation><orgName type="institution">Drug Discovery and Design Center</orgName><orgName type="laboratory">State Key Laboratory of Drug Research</orgName><orgName type="institution">Shanghai Institute of Materia Medica</orgName><orgName type="institution">Shanghai Institutes for Biological Sciences</orgName><orgName type="department">Graduate School</orgName><orgName type="institution">Chinese Academy of Sciences</orgName><address><addrLine>Shanghai 201203</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>Shanghai Institute of Plant Physiology and Ecology</addrLine><addrLine>Shanghai Institutes for Biological Sciences</addrLine><addrLine>Graduate School</addrLine><addrLine>Chinese Academy of Sciences</addrLine><addrLine>300 Fenglin Road</addrLine><addrLine>Shanghai 200032</addrLine><country key="CN" xml:lang="en">CHINA</country><addrLine>and School of Pharmacy</addrLine><addrLine>East-China University of Science and Technology</addrLine><addrLine>Shanghai 200237</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note></author><author xml:id="author-0009" role="corresp"><persName><surname>Shen</surname><forename type="first">Xu</forename></persName><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note><affiliation role="corresp"> To whom correspondence should be addressed. E-mail: xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.). Telephone: 86-21-50807188. Fax: 86-21-50807088.</affiliation></author><author xml:id="author-0010" role="corresp"><persName><surname>Jiang</surname><forename type="first">Hualiang</forename></persName><note place="foot" n="bi0482396AF3"><ref>‡</ref><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></note><note place="foot" n="bi0482396AF6"><ref>⊥</ref><p>
East-China University of Science and Technology.</p></note><affiliation role="corresp"> To whom correspondence should be addressed. E-mail: xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.). Telephone: 86-21-50807188. Fax: 86-21-50807088.</affiliation></author><idno type="istex">1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052</idno><idno type="ark">ark:/67375/TPS-NZ5V5XNP-C</idno><idno type="DOI">10.1021/bi0482396</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">2005</date><date type="published">2005</date><biblScope unit="vol">44</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="1453">1453</biblScope><biblScope unit="page" to="1463">1463</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="bi0482396n00001.tif"></graphic></p><p>Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus
(SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is
believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We
have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of
SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques.
For the thermally induced unfolding at pH 7.4, the apparent activation energy (<hi rend="italic">E</hi><hi rend="subscript">app</hi>) and transition midpoint
temperature (<hi rend="italic">T</hi><hi rend="subscript">m</hi>) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD
spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a
relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent
spectra resulted in consistent values of the transition middle concentration of the denaturant (<hi rend="italic">C</hi><hi rend="subscript">m</hi>, ranging
from 2.30 to 2.45 M) and the standard free energy change (Δ<hi rend="italic">G</hi>°, ranging from 2.1 to 2.5 kcal/mol) for
the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding
of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding
reaction rates and relaxation times were determined at various GuHCl concentrations, and the <hi rend="italic">C</hi><hi rend="subscript">m</hi> value
was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations.
Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b
folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the
protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional
model was also used to explain the recently published experimental results of S1−ACE-2 binding and
immunizations.
</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/bi0482396</article-id><article-categories><subj-group subj-group-type="document-type-name"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment
(SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with
Three-Dimensional Structural Modeling<xref rid="bi0482396AF2">†</xref></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Yu</surname><given-names>Changying</given-names></name><xref rid="bi0482396AF3">‡</xref><xref rid="bi0482396AF4">§</xref></contrib><contrib contrib-type="author"><name><surname>Gui</surname><given-names>Chunshan</given-names></name><xref rid="bi0482396AF3">‡</xref><xref rid="bi0482396AF4">§</xref></contrib><contrib contrib-type="author"><name><surname>Luo</surname><given-names>Haibin</given-names></name><xref rid="bi0482396AF3">‡</xref></contrib><contrib contrib-type="author"><name><surname>Chen</surname><given-names>Lili</given-names></name><xref rid="bi0482396AF3">‡</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Liang</given-names></name><xref rid="bi0482396AF3">‡</xref></contrib><contrib contrib-type="author"><name><surname>Yu</surname><given-names>Hao</given-names></name><xref rid="bi0482396AF5">‖</xref></contrib><contrib contrib-type="author"><name><surname>Yang</surname><given-names>Sheng</given-names></name><xref rid="bi0482396AF5">‖</xref></contrib><contrib contrib-type="author"><name><surname>Jiang</surname><given-names>Weihong</given-names></name><xref rid="bi0482396AF5">‖</xref></contrib><contrib contrib-type="author"><name><surname>Shen</surname><given-names>Jianhua</given-names></name><xref rid="bi0482396AF3">‡</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Shen</surname><given-names>Xu</given-names></name><xref rid="bi0482396AF1">*</xref><xref rid="bi0482396AF3">‡</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Jiang</surname><given-names>Hualiang</given-names></name><xref rid="bi0482396AF1">*</xref><xref rid="bi0482396AF3">‡</xref><xref rid="bi0482396AF6">⊥</xref></contrib><aff>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai
Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Institute
of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,
300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,
Shanghai 200237, China
</aff></contrib-group><author-notes><fn id="bi0482396AF3"><label>‡</label><p>
Shanghai Institute of Materia Medica, Shanghai Institutes for
Biological Sciences, Graduate School, Chinese Academy of Sciences.
</p></fn><fn id="bi0482396AF4"><label>§</label><p>
These authors contributed equally to this work.</p></fn><fn id="bi0482396AF5"><label>‖</label><p>
Shanghai Institute of Plant Physiology & Ecology, Shanghai
Institutes for Biological Sciences, Graduate School, Chinese Academy
of Sciences.</p></fn><corresp id="bi0482396AF1">
To whom correspondence should be addressed. E-mail:
xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).
Telephone: 86-21-50807188. Fax: 86-21-50807088.</corresp><fn id="bi0482396AF6"><label>⊥</label><p>
East-China University of Science and Technology.</p></fn></author-notes><pub-date pub-type="epub"><day>6</day><month>01</month><year>2005</year></pub-date><pub-date pub-type="ppub"><day>08</day><month>02</month><year>2005</year></pub-date><volume>44</volume><issue>5</issue><fpage>1453</fpage><lpage>1463</lpage><supplementary-material xlink:href="bi0482396si20041113_084007.pdf"></supplementary-material><history><date date-type="received"><day>14</day><month>08</month><year>2004</year></date><date date-type="rev-recd"><day>13</day><month>11</month><year>2004</year></date><date date-type="asap"><day>6</day><month>01</month><year>2005</year></date><date date-type="issue-pub"><day>08</day><month>02</month><year>2005</year></date></history><permissions><copyright-statement>Copyright © 2005 American Chemical Society</copyright-statement><copyright-year>2005</copyright-year><copyright-holder>American Chemical Society</copyright-holder></permissions><abstract><graphic content-type="abstract-graphic" xlink:href="bi0482396n00001.tif"></graphic><p>Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus
(SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is
believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We
have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of
SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques.
For the thermally induced unfolding at pH 7.4, the apparent activation energy (<italic>E</italic><sub>app</sub>) and transition midpoint
temperature (<italic>T</italic><sub>m</sub>) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD
spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a
relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent
spectra resulted in consistent values of the transition middle concentration of the denaturant (<italic>C</italic><sub>m</sub>, ranging
from 2.30 to 2.45 M) and the standard free energy change (Δ<italic>G</italic>°, ranging from 2.1 to 2.5 kcal/mol) for
the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding
of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding
reaction rates and relaxation times were determined at various GuHCl concentrations, and the <italic>C</italic><sub>m</sub> value
was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations.
Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b
folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the
protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional
model was also used to explain the recently published experimental results of S1−ACE-2 binding and
immunizations.
</p></abstract><custom-meta-group><custom-meta><meta-name>document-id-old-9</meta-name><meta-value>bi0482396</meta-value></custom-meta></custom-meta-group></article-meta><notes id="bi0482396AF2"><label>†</label><p>
This work was supported by the State Key Program of Basic
Research of China (Grants 2003CB514125, 2002CB512801, 2002CB512802, and 2002CB512807), the National Natural Science Foundation
of China (Grants 20372069 and 20472095), Shanghai Basic Research
Project from the Shanghai Science and Technology Commission (Grants
02DJ14070, 03DZ19228, and 03DZ19212), the 863 Hi-Tech Program
(Grants 2001AA235051, 2002CB512807, and 2002AA233011), the
Sino-European Project on SARS Diagnostics and Antivirals (Proposal/Contract 003831), and the special programs of oppugning SARS from
the Ministry of Science and Technology, Chinese Academy of Sciences,
the National Natural Science Foundation of China, the Sino-German
Science Center, and the Shanghai Science and Technology Commission.
</p></notes></front><body></body><back></back></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†</title></titleInfo><titleInfo contentType="CDATA"><title>Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†</title></titleInfo><name type="personal"><namePart type="family">YU</namePart><namePart type="given">Changying</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><affiliation> These authors contributed equally to this work.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">GUI</namePart><namePart type="given">Chunshan</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><affiliation> These authors contributed equally to this work.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">LUO</namePart><namePart type="given">Haibin</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">CHEN</namePart><namePart type="given">Lili</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">ZHANG</namePart><namePart type="given">Liang</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">YU</namePart><namePart type="given">Hao</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">YANG</namePart><namePart type="given">Sheng</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">JIANG</namePart><namePart type="given">Weihong</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Plant Physiology & Ecology, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academyof Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="family">SHEN</namePart><namePart type="given">Jianhua</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal" displayLabel="corresp"><namePart type="family">SHEN</namePart><namePart type="given">Xu</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal" displayLabel="corresp"><namePart type="family">JIANG</namePart><namePart type="given">Hualiang</namePart><affiliation>Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, ShanghaiInstitutes for Biological Sciences, Graduate School, Chinese Academy of Sciences, Shanghai 201203, China, Shanghai Instituteof Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Graduate School, Chinese Academy of Sciences,300 Fenglin Road, Shanghai 200032, China, and School of Pharmacy, East-China University of Science and Technology,Shanghai 200237, China</affiliation><affiliation> Shanghai Institute of Materia Medica, Shanghai Institutes forBiological Sciences, Graduate School, Chinese Academy of Sciences.</affiliation><affiliation> East-China University of Science and Technology.</affiliation><affiliation> To whom correspondence should be addressed. E-mail:xshen@mail.shcnc.ac.cn (X.S.) or hljiang@mail.shcnc.ac.cn (H.J.).Telephone: 86-21-50807188. Fax: 86-21-50807088.</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">2005-01-06</dateCreated><dateIssued encoding="w3cdtf">2005-02-08</dateIssued><copyrightDate encoding="w3cdtf">2005</copyrightDate></originInfo><note type="footnote" ID="bi0482396AF2"> This work was supported by the State Key Program of Basic Research of China (Grants 2003CB514125, 2002CB512801, 2002CB512802, and 2002CB512807), the National Natural Science Foundation of China (Grants 20372069 and 20472095), Shanghai Basic Research Project from the Shanghai Science and Technology Commission (Grants 02DJ14070, 03DZ19228, and 03DZ19212), the 863 Hi-Tech Program (Grants 2001AA235051, 2002CB512807, and 2002AA233011), the Sino-European Project on SARS Diagnostics and Antivirals (Proposal/Contract 003831), and the special programs of oppugning SARS from the Ministry of Science and Technology, Chinese Academy of Sciences, the National Natural Science Foundation of China, the Sino-German Science Center, and the Shanghai Science and Technology Commission.</note><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract>Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus (SARS_CoV) infection. The immunological fragment of the S protein (Ala251−His641, SARS_S1b) is believed to be essential for SARS_CoV entering the host cell through S protein−ACE-2 interaction. We have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques. For the thermally induced unfolding at pH 7.4, the apparent activation energy (Eapp) and transition midpoint temperature (Tm) were determined to be 16.3 ± 0.2 kcal/mol and 52.5 ± 0.4 °C, respectively. The CD spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent spectra resulted in consistent values of the transition middle concentration of the denaturant (Cm, ranging from 2.30 to 2.45 M) and the standard free energy change (ΔG°, ranging from 2.1 to 2.5 kcal/mol) for the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding reaction rates and relaxation times were determined at various GuHCl concentrations, and the Cm value was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations. Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b folded as a globular-like structure by β-sheets and loops; two of the four tryptophans are located on the protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional model was also used to explain the recently published experimental results of S1−ACE-2 binding and immunizations.</abstract><note type="footnote" ID="bi0482396AF2"> This work was supported by the State Key Program of Basic Research of China (Grants 2003CB514125, 2002CB512801, 2002CB512802, and 2002CB512807), the National Natural Science Foundation of China (Grants 20372069 and 20472095), Shanghai Basic Research Project from the Shanghai Science and Technology Commission (Grants 02DJ14070, 03DZ19228, and 03DZ19212), the 863 Hi-Tech Program (Grants 2001AA235051, 2002CB512807, and 2002AA233011), the Sino-European Project on SARS Diagnostics and Antivirals (Proposal/Contract 003831), and the special programs of oppugning SARS from the Ministry of Science and Technology, Chinese Academy of Sciences, the National Natural Science Foundation of China, the Sino-German Science Center, and the Shanghai Science and Technology Commission.</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>2005</date><detail type="volume"><caption>vol.</caption><number>44</number></detail><detail type="issue"><caption>no.</caption><number>5</number></detail><extent unit="pages"><start>1453</start><end>1463</end></extent></part></relatedItem><identifier type="istex">1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052</identifier><identifier type="ark">ark:/67375/TPS-NZ5V5XNP-C</identifier><identifier type="DOI">10.1021/bi0482396</identifier><accessCondition type="use and reproduction" contentType="restricted">Copyright © 2005 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-NZ5V5XNP-C/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/1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052/annexes/tiff</uri></json:item><json:item><extension>gif</extension><original>true</original><mimetype>image/gif</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-NZ5V5XNP-C/annexes.gif</uri></json:item><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/TPS-NZ5V5XNP-C/annexes.pdf</uri></json:item></annexes><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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|étape= Corpus
|type= RBID
|clé= ISTEX:1B3B857EECDEE18BE6E04BE4F62B875F5C8F7052
|texte= Folding of the SARS Coronavirus Spike Glycoprotein Immunological Fragment (SARS_S1B): Thermodynamic and Kinetic Investigation Correlating with Three-Dimensional Structural Modeling†
}}
| This area was generated with Dilib version V0.6.33. |  |