X-ray Structure and Enzymatic Activity Profile of a Core Papain-like Protease of MERS Coronavirus with utility for structure-based drug design
Identifieur interne : 000B18 ( Main/Exploration ); précédent : 000B17; suivant : 000B19X-ray Structure and Enzymatic Activity Profile of a Core Papain-like Protease of MERS Coronavirus with utility for structure-based drug design
Auteurs : Jozlyn R. Clasman [États-Unis] ; Yahira M. Báez-Santos [États-Unis] ; Robert C. Mettelman [États-Unis] ; Amornrat O Rien [États-Unis] ; Susan C. Baker [États-Unis] ; Andrew D. Mesecar [États-Unis]Source :
- Scientific Reports [ 2045-2322 ] ; 2017.
Descripteurs français
- KwdFr :
- Cinétique, Conception de médicament, Coronavirus du syndrome respiratoire du Moyen-Orient (enzymologie), Cristallographie aux rayons X, Domaines protéiques, Humains, Luciferases (métabolisme), Maturation post-traductionnelle des protéines, Papaïne (), Polyprotéines (), Stabilité enzymatique, Techniques de biocapteur, Température, Transfert d'énergie par résonance de fluorescence, Ubiquitine ().
- MESH :
- enzymologie : Coronavirus du syndrome respiratoire du Moyen-Orient.
- métabolisme : Luciferases.
- Cinétique, Conception de médicament, Cristallographie aux rayons X, Domaines protéiques, Humains, Maturation post-traductionnelle des protéines, Papaïne, Polyprotéines, Stabilité enzymatique, Techniques de biocapteur, Température, Transfert d'énergie par résonance de fluorescence, Ubiquitine.
English descriptors
- KwdEn :
- Biosensing Techniques, Crystallography, X-Ray, Drug Design, Enzyme Stability, Fluorescence Resonance Energy Transfer, Humans, Kinetics, Luciferases (metabolism), Middle East Respiratory Syndrome Coronavirus (enzymology), Papain (chemistry), Polyproteins (chemistry), Protein Domains, Protein Processing, Post-Translational, Temperature, Ubiquitin (chemistry).
- MESH :
- chemical , chemistry : Papain, Polyproteins, Ubiquitin.
- chemical , metabolism : Luciferases.
- enzymology : Middle East Respiratory Syndrome Coronavirus.
- Biosensing Techniques, Crystallography, X-Ray, Drug Design, Enzyme Stability, Fluorescence Resonance Energy Transfer, Humans, Kinetics, Protein Domains, Protein Processing, Post-Translational, Temperature.
Abstract
Ubiquitin-like domain 2 (Ubl2) is immediately adjacent to the N-terminus of the papain-like protease (PLpro) domain in coronavirus polyproteins, and it may play a critical role in protease regulation and stability as well as in viral infection. However, our recent cellular studies reveal that removing the Ubl2 domain from MERS PLpro has no effect on its ability to process the viral polyprotein or act as an interferon antagonist, which involves deubiquitinating and deISGylating cellular proteins. Here, we test the hypothesis that the Ubl2 domain is not required for the catalytic function of MERS PLpro
The online version of this article (doi:10.1038/srep40292) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1038/srep40292
PubMed: 28079137
PubMed Central: 5228125
Affiliations:
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However, our recent cellular studies reveal that removing the Ubl2 domain from MERS PLpro has no effect on its ability to process the viral polyprotein or act as an interferon antagonist, which involves deubiquitinating and deISGylating cellular proteins. Here, we test the hypothesis that the Ubl2 domain is not required for the catalytic function of MERS PLpro <italic>in vitro</italic>. The X-ray structure of MERS PLpro-∆Ubl2 was determined to 1.9 Å and compared to PLpro containing the N-terminal Ubl2 domain. While the structures were nearly identical, the PLpro-∆Ubl2 enzyme revealed the intact structure of the substrate-binding loop. Moreover, PLpro-∆Ubl2 catalysis against different substrates and a purported inhibitor revealed no differences in catalytic efficiency, substrate specificity, and inhibition. Further, no changes in thermal stability were observed between enzymes. We conclude that the catalytic core of MERS PLpro, i.e. without the Ubl2 domain, is sufficient for catalysis and stability <italic>in vitro</italic> with utility to evaluate potential inhibitors as a platform for structure-based drug design.</p><sec><title>Supplementary information</title><p>The online version of this article (doi:10.1038/srep40292) contains supplementary material, which is available to authorized users.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Peiris, Js" uniqKey="Peiris J">JS Peiris</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y Guan</name></author><author><name sortKey="Yuen, Ky" uniqKey="Yuen K">KY Yuen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chan, Jf" uniqKey="Chan J">JF Chan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Barretto, N" uniqKey="Barretto N">N 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sortKey="Mielech, Am" uniqKey="Mielech A">AM Mielech</name></author><author><name sortKey="Deng, X" uniqKey="Deng X">X Deng</name></author><author><name sortKey="Baker, Sc" uniqKey="Baker S">SC Baker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bekes, M" uniqKey="Bekes M">M Békés</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhu, X" uniqKey="Zhu X">X Zhu</name></author><author><name sortKey="Menard, R" uniqKey="Menard R">R Ménard</name></author><author><name sortKey="Sulea, T" uniqKey="Sulea T">T Sulea</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Giner Sorolla, A" uniqKey="Giner Sorolla A">A Giner-Sorolla</name></author><author><name sortKey="Bendich, A" uniqKey="Bendich A">A Bendich</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Medebielle, M" uniqKey="Medebielle M">M Medebielle</name></author><author><name sortKey="Fujii, S" uniqKey="Fujii S">S Fujii</name></author><author><name sortKey="Kato, K" uniqKey="Kato K">K Kato</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, H" uniqKey="Lee H">H Lee</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Baell, J" uniqKey="Baell J">J Baell</name></author><author><name sortKey="Walters, Ma" uniqKey="Walters M">MA Walters</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Otwinowski, Z" uniqKey="Otwinowski Z">Z Otwinowski</name></author><author><name sortKey="Minor, W" uniqKey="Minor W">W Minor</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Emsley, P" uniqKey="Emsley P">P Emsley</name></author><author><name sortKey="Cowtan, K" uniqKey="Cowtan K">K Cowtan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Afonine, Pv" uniqKey="Afonine P">PV Afonine</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Illinois</li><li>Indiana</li></region></list><tree><country name="États-Unis"><region name="Indiana"><name sortKey="Clasman, Jozlyn R" sort="Clasman, Jozlyn R" uniqKey="Clasman J" first="Jozlyn R." last="Clasman">Jozlyn R. Clasman</name></region><name sortKey="Baez Santos, Yahira M" sort="Baez Santos, Yahira M" uniqKey="Baez Santos Y" first="Yahira M." last="Báez-Santos">Yahira M. Báez-Santos</name><name sortKey="Baker, Susan C" sort="Baker, Susan C" uniqKey="Baker S" first="Susan C." last="Baker">Susan C. Baker</name><name sortKey="Mesecar, Andrew D" sort="Mesecar, Andrew D" uniqKey="Mesecar A" first="Andrew D." last="Mesecar">Andrew D. Mesecar</name><name sortKey="Mesecar, Andrew D" sort="Mesecar, Andrew D" uniqKey="Mesecar A" first="Andrew D." last="Mesecar">Andrew D. Mesecar</name><name sortKey="Mesecar, Andrew D" sort="Mesecar, Andrew D" uniqKey="Mesecar A" first="Andrew D." last="Mesecar">Andrew D. Mesecar</name><name sortKey="Mettelman, Robert C" sort="Mettelman, Robert C" uniqKey="Mettelman R" first="Robert C." last="Mettelman">Robert C. Mettelman</name><name sortKey="O Rien, Amornrat" sort="O Rien, Amornrat" uniqKey="O Rien A" first="Amornrat" last="O Rien">Amornrat O Rien</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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