MERS-CoV papain-like protease (PLpro): expression, purification, and spectroscopic/thermodynamic characterization.
Identifieur interne : 000A99 ( PubMed/Checkpoint ); précédent : 000A98; suivant : 000B00MERS-CoV papain-like protease (PLpro): expression, purification, and spectroscopic/thermodynamic characterization.
Auteurs : Ajamaluddin Malik [Arabie saoudite] ; Mohammad A. Alsenaidy [Arabie saoudite]Source :
- 3 Biotech [ 2190-572X ] ; 2017.
Abstract
Within a decade, MERS-CoV emerged with nearly four times higher case fatality rate than an earlier outbreak of SARS-CoV and spread out in 27 countries in short span of time. As an emerging virus, combating it requires an in-depth understanding of its molecular machinery. Therefore, conformational characterization studies of coronavirus proteins are necessary to advance our knowledge of the matter for the development of antiviral therapies. In this study, MERS-CoV papain-like protease (PLpro) was recombinantly expressed and purified. Thermal folding pathway and thermodynamic properties were characterized using dynamic multimode spectroscopy (DMS) and thermal shift assay. DMS study showed that the PLpro undergoes a single thermal transition and follows a pathway of two-state folding with T m and van't Hoff enthalpy values of 54.4 ± 0.1 °C and 317.1 ± 3.9 kJ/mol, respectively. An orthogonal technique based on intrinsic tryptophan fluorescence also showed that MERS-CoV PLpro undergoes a single thermal transition and unfolds via a pathway of two-state folding with a T m value of 51.4 °C. Our findings provide significant understandings of the thermodynamic and structural properties of MERS-CoV PLpro.
DOI: 10.1007/s13205-017-0744-3
PubMed: 28560640
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">MERS-CoV papain-like protease (PL<sup>pro</sup>): expression, purification, and spectroscopic/thermodynamic characterization.</title><author><name sortKey="Malik, Ajamaluddin" sort="Malik, Ajamaluddin" uniqKey="Malik A" first="Ajamaluddin" last="Malik">Ajamaluddin Malik</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Protein Research Chair, College of Science, King Saud University, PO Box 2455, Riyadh, 11451, Saudi Arabia. amalik@ksu.edu.sa.</nlm:affiliation><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Department of Biochemistry, Protein Research Chair, College of Science, King Saud University, PO Box 2455, Riyadh, 11451</wicri:regionArea><wicri:noRegion>11451</wicri:noRegion></affiliation></author><author><name sortKey="Alsenaidy, Mohammad A" sort="Alsenaidy, Mohammad A" uniqKey="Alsenaidy M" first="Mohammad A" last="Alsenaidy">Mohammad A. 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Alsenaidy</name><affiliation wicri:level="1"><nlm:affiliation>Vaccines and Biologics Research Unit, Department of Pharmaceutics, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Saudi Arabia.</nlm:affiliation><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Vaccines and Biologics Research Unit, Department of Pharmaceutics, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451</wicri:regionArea><wicri:noRegion>11451</wicri:noRegion></affiliation></author></analytic><series><title level="j">3 Biotech</title><idno type="ISSN">2190-572X</idno><imprint><date when="2017" type="published">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Within a decade, MERS-CoV emerged with nearly four times higher case fatality rate than an earlier outbreak of SARS-CoV and spread out in 27 countries in short span of time. As an emerging virus, combating it requires an in-depth understanding of its molecular machinery. Therefore, conformational characterization studies of coronavirus proteins are necessary to advance our knowledge of the matter for the development of antiviral therapies. In this study, MERS-CoV papain-like protease (PL<sup>pro</sup>) was recombinantly expressed and purified. Thermal folding pathway and thermodynamic properties were characterized using dynamic multimode spectroscopy (DMS) and thermal shift assay. DMS study showed that the PL<sup>pro</sup> undergoes a single thermal transition and follows a pathway of two-state folding with T <sub>m</sub> and van't Hoff enthalpy values of 54.4 ± 0.1 °C and 317.1 ± 3.9 kJ/mol, respectively. An orthogonal technique based on intrinsic tryptophan fluorescence also showed that MERS-CoV PL<sup>pro</sup> undergoes a single thermal transition and unfolds via a pathway of two-state folding with a T <sub>m</sub> value of 51.4 °C. Our findings provide significant understandings of the thermodynamic and structural properties of MERS-CoV PL<sup>pro</sup>.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">28560640</PMID><DateRevised><Year>2019</Year><Month>11</Month><Day>20</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">2190-572X</ISSN><JournalIssue CitedMedium="Print"><Volume>7</Volume><Issue>2</Issue><PubDate><Year>2017</Year><Month>Jun</Month></PubDate></JournalIssue><Title>3 Biotech</Title><ISOAbbreviation>3 Biotech</ISOAbbreviation></Journal><ArticleTitle>MERS-CoV papain-like protease (PL<sup>pro</sup>): expression, purification, and spectroscopic/thermodynamic characterization.</ArticleTitle><Pagination><MedlinePgn>100</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s13205-017-0744-3</ELocationID><Abstract><AbstractText>Within a decade, MERS-CoV emerged with nearly four times higher case fatality rate than an earlier outbreak of SARS-CoV and spread out in 27 countries in short span of time. As an emerging virus, combating it requires an in-depth understanding of its molecular machinery. Therefore, conformational characterization studies of coronavirus proteins are necessary to advance our knowledge of the matter for the development of antiviral therapies. In this study, MERS-CoV papain-like protease (PL<sup>pro</sup>) was recombinantly expressed and purified. Thermal folding pathway and thermodynamic properties were characterized using dynamic multimode spectroscopy (DMS) and thermal shift assay. DMS study showed that the PL<sup>pro</sup> undergoes a single thermal transition and follows a pathway of two-state folding with T <sub>m</sub> and van't Hoff enthalpy values of 54.4 ± 0.1 °C and 317.1 ± 3.9 kJ/mol, respectively. An orthogonal technique based on intrinsic tryptophan fluorescence also showed that MERS-CoV PL<sup>pro</sup> undergoes a single thermal transition and unfolds via a pathway of two-state folding with a T <sub>m</sub> value of 51.4 °C. Our findings provide significant understandings of the thermodynamic and structural properties of MERS-CoV PL<sup>pro</sup>.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Malik</LastName><ForeName>Ajamaluddin</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Protein Research Chair, College of Science, King Saud University, PO Box 2455, Riyadh, 11451, Saudi Arabia. amalik@ksu.edu.sa.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Alsenaidy</LastName><ForeName>Mohammad A</ForeName><Initials>MA</Initials><AffiliationInfo><Affiliation>Vaccines and Biologics Research Unit, Department of Pharmaceutics, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Saudi Arabia.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate 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IdType="pubmed">25395330</ArticleId></ArticleIdList></Reference><Reference><Citation>Front Med. 2016 Jun;10 (2):111-9</Citation><ArticleIdList><ArticleId IdType="pubmed">27146399</ArticleId></ArticleIdList></Reference><Reference><Citation>J Tradit Chin Med. 2003 Sep;23(3):214-7</Citation><ArticleIdList><ArticleId IdType="pubmed">14535196</ArticleId></ArticleIdList></Reference><Reference><Citation>Respirology. 2003 Nov;8 Suppl:S36-40</Citation><ArticleIdList><ArticleId IdType="pubmed">15018132</ArticleId></ArticleIdList></Reference><Reference><Citation>Int J Biol Macromol. 2016 Jul;88:313-9</Citation><ArticleIdList><ArticleId IdType="pubmed">27044344</ArticleId></ArticleIdList></Reference><Reference><Citation>Eur Biophys J. 2015 Dec;44(8):667-75</Citation><ArticleIdList><ArticleId IdType="pubmed">26219412</ArticleId></ArticleIdList></Reference><Reference><Citation>FEBS J. 2014 Sep;281(18):4085-96</Citation><ArticleIdList><ArticleId IdType="pubmed">25039866</ArticleId></ArticleIdList></Reference><Reference><Citation>Int J Pharm. 2005 Apr 27;294(1-2):193-9</Citation><ArticleIdList><ArticleId IdType="pubmed">15814244</ArticleId></ArticleIdList></Reference><Reference><Citation>Chest. 2005 Sep;128(3):1393-400</Citation><ArticleIdList><ArticleId IdType="pubmed">16162734</ArticleId></ArticleIdList></Reference><Reference><Citation>Antiviral Res. 2014 Sep;109:72-82</Citation><ArticleIdList><ArticleId IdType="pubmed">24992731</ArticleId></ArticleIdList></Reference><Reference><Citation>Emerg Infect Dis. 2014 Jul;20(7):1223-6</Citation><ArticleIdList><ArticleId IdType="pubmed">24960370</ArticleId></ArticleIdList></Reference><Reference><Citation>Virol J. 2015 Dec 22;12 :209</Citation><ArticleIdList><ArticleId IdType="pubmed">26690088</ArticleId></ArticleIdList></Reference><Reference><Citation>Virus Res. 2012 Apr;165(1):112-7</Citation><ArticleIdList><ArticleId IdType="pubmed">22349148</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Protoc. 2006;1(6):2527-35</Citation><ArticleIdList><ArticleId IdType="pubmed">17406506</ArticleId></ArticleIdList></Reference><Reference><Citation>J Gen Virol. 2014 Mar;95(Pt 3):614-26</Citation><ArticleIdList><ArticleId IdType="pubmed">24362959</ArticleId></ArticleIdList></Reference><Reference><Citation>Photosynth Res. 2015 Dec;126(2-3):427-35</Citation><ArticleIdList><ArticleId IdType="pubmed">26071019</ArticleId></ArticleIdList></Reference><Reference><Citation>Clin Endocrinol (Oxf). 2005 Aug;63(2):197-202</Citation><ArticleIdList><ArticleId IdType="pubmed">16060914</ArticleId></ArticleIdList></Reference><Reference><Citation>J Korean Med Sci. 2016 Apr;31(4):635-40</Citation><ArticleIdList><ArticleId IdType="pubmed">27051251</ArticleId></ArticleIdList></Reference><Reference><Citation>Int J Mol Sci. 2014 Dec 05;15(12):22518-38</Citation><ArticleIdList><ArticleId IdType="pubmed">25490136</ArticleId></ArticleIdList></Reference><Reference><Citation>ACS Nano. 2012 Oct 23;6(10):9335-46</Citation><ArticleIdList><ArticleId IdType="pubmed">22857653</ArticleId></ArticleIdList></Reference><Reference><Citation>RNA Biol. 2011 Mar-Apr;8(2):270-9</Citation><ArticleIdList><ArticleId IdType="pubmed">21593585</ArticleId></ArticleIdList></Reference><Reference><Citation>Biophys Chem. 1991 Nov;41(2):185-91</Citation><ArticleIdList><ArticleId IdType="pubmed">1773011</ArticleId></ArticleIdList></Reference><Reference><Citation>J Virol. 2015 May;89(9):4907-17</Citation><ArticleIdList><ArticleId IdType="pubmed">25694594</ArticleId></ArticleIdList></Reference><Reference><Citation>Travel Med Infect Dis. 2014 Sep-Oct;12 (5):422-8</Citation><ArticleIdList><ArticleId IdType="pubmed">25047726</ArticleId></ArticleIdList></Reference><Reference><Citation>J Virol. 2013 Nov;87(21):11955-62</Citation><ArticleIdList><ArticleId IdType="pubmed">23986593</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biomed Sci. 2014 Jun 04;21:54</Citation><ArticleIdList><ArticleId IdType="pubmed">24898546</ArticleId></ArticleIdList></Reference><Reference><Citation>Health Expect. 2016 Jun;19(3):762-72</Citation><ArticleIdList><ArticleId IdType="pubmed">25220505</ArticleId></ArticleIdList></Reference><Reference><Citation>Pediatr Pulmonol. 2007 Apr;42(4):393-6</Citation><ArticleIdList><ArticleId IdType="pubmed">17352399</ArticleId></ArticleIdList></Reference><Reference><Citation>J Virol. 2010 May;84(9):4619-29</Citation><ArticleIdList><ArticleId IdType="pubmed">20181693</ArticleId></ArticleIdList></Reference><Reference><Citation>J Vet Med Sci. 2016 Mar;78(3):355-63</Citation><ArticleIdList><ArticleId IdType="pubmed">26537549</ArticleId></ArticleIdList></Reference><Reference><Citation>Protein Sci. 2000 Jul;9(7):1416-9</Citation><ArticleIdList><ArticleId IdType="pubmed">10933511</ArticleId></ArticleIdList></Reference><Reference><Citation>Eur J Biochem. 2003 Feb;270(4):745-56</Citation><ArticleIdList><ArticleId IdType="pubmed">12581214</ArticleId></ArticleIdList></Reference><Reference><Citation>Viruses. 2012 Apr;4(4):637-53</Citation><ArticleIdList><ArticleId IdType="pubmed">22590689</ArticleId></ArticleIdList></Reference><Reference><Citation>Cell Res. 2008 Nov;18(11):1105-13</Citation><ArticleIdList><ArticleId IdType="pubmed">18957937</ArticleId></ArticleIdList></Reference><Reference><Citation>Biophys J. 2006 May 15;90(10):3704-11</Citation><ArticleIdList><ArticleId IdType="pubmed">16500981</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Arabie saoudite</li></country></list><tree><country name="Arabie saoudite"><noRegion><name sortKey="Malik, Ajamaluddin" sort="Malik, Ajamaluddin" uniqKey="Malik A" first="Ajamaluddin" last="Malik">Ajamaluddin Malik</name></noRegion><name sortKey="Alsenaidy, Mohammad A" sort="Alsenaidy, Mohammad A" uniqKey="Alsenaidy M" first="Mohammad A" last="Alsenaidy">Mohammad A. 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