2hob
From Proteopedia
(New page: 200px<br /><applet load="2hob" size="450" color="white" frame="true" align="right" spinBox="true" caption="2hob, resolution 1.95Å" /> '''Crystal structure of...) |
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| - | [[Image:2hob.jpg|left|200px]]<br /><applet load="2hob" size=" | + | [[Image:2hob.jpg|left|200px]]<br /><applet load="2hob" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2hob, resolution 1.95Å" /> | caption="2hob, resolution 1.95Å" /> | ||
'''Crystal structure of SARS-CoV main protease with authentic N and C-termini in complex with a Michael acceptor N3'''<br /> | '''Crystal structure of SARS-CoV main protease with authentic N and C-termini in complex with a Michael acceptor N3'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The viral proteases have proven to be the most selective and useful for | + | The viral proteases have proven to be the most selective and useful for removing the fusion tags in fusion protein expression systems. As a key enzyme in the viral life-cycle, the main protease (M(pro)) is most attractive for drug design targeting the SARS coronavirus (SARS-CoV), the etiological agent responsible for the outbreak of severe acute respiratory syndrome (SARS) in 2003. In this study, SARS-CoV M(pro) was used to specifically remove the GST tag in a new fusion protein expression system. We report a new method to produce wild-type (WT) SARS-CoV M(pro) with authentic N and C termini, and compare the activity of WT protease with those of three different types of SARS-CoV M(pro) with additional residues at the N or C terminus. Our results show that additional residues at the N terminus, but not at the C terminus, of M(pro) are detrimental to enzyme activity. To explain this, the crystal structures of WT SARS-CoV M(pro) and its complex with a Michael acceptor inhibitor were determined to 1.6 Angstroms and 1.95 Angstroms resolution respectively. These crystal structures reveal that the first residue of this protease is important for sustaining the substrate-binding pocket and inhibitor binding. This study suggests that SARS-CoV M(pro) could serve as a new tag-cleavage endopeptidase for protein overproduction, and the WT SARS-CoV M(pro) is more appropriate for mechanistic characterization and inhibitor design. |
==About this Structure== | ==About this Structure== | ||
| - | 2HOB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sars_coronavirus Sars coronavirus] with 3IH as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 2HOB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sars_coronavirus Sars coronavirus] with <scene name='pdbligand=3IH:'>3IH</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2HOB OCA]. |
==Reference== | ==Reference== | ||
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[[Category: sars-cov]] | [[Category: sars-cov]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:43:54 2008'' |
Revision as of 15:43, 21 February 2008
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Crystal structure of SARS-CoV main protease with authentic N and C-termini in complex with a Michael acceptor N3
Overview
The viral proteases have proven to be the most selective and useful for removing the fusion tags in fusion protein expression systems. As a key enzyme in the viral life-cycle, the main protease (M(pro)) is most attractive for drug design targeting the SARS coronavirus (SARS-CoV), the etiological agent responsible for the outbreak of severe acute respiratory syndrome (SARS) in 2003. In this study, SARS-CoV M(pro) was used to specifically remove the GST tag in a new fusion protein expression system. We report a new method to produce wild-type (WT) SARS-CoV M(pro) with authentic N and C termini, and compare the activity of WT protease with those of three different types of SARS-CoV M(pro) with additional residues at the N or C terminus. Our results show that additional residues at the N terminus, but not at the C terminus, of M(pro) are detrimental to enzyme activity. To explain this, the crystal structures of WT SARS-CoV M(pro) and its complex with a Michael acceptor inhibitor were determined to 1.6 Angstroms and 1.95 Angstroms resolution respectively. These crystal structures reveal that the first residue of this protease is important for sustaining the substrate-binding pocket and inhibitor binding. This study suggests that SARS-CoV M(pro) could serve as a new tag-cleavage endopeptidase for protein overproduction, and the WT SARS-CoV M(pro) is more appropriate for mechanistic characterization and inhibitor design.
About this Structure
2HOB is a Single protein structure of sequence from Sars coronavirus with as ligand. Full crystallographic information is available from OCA.
Reference
Production of authentic SARS-CoV M(pro) with enhanced activity: application as a novel tag-cleavage endopeptidase for protein overproduction., Xue X, Yang H, Shen W, Zhao Q, Li J, Yang K, Chen C, Jin Y, Bartlam M, Rao Z, J Mol Biol. 2007 Feb 23;366(3):965-75. Epub 2006 Dec 1. PMID:17189639
Page seeded by OCA on Thu Feb 21 17:43:54 2008
Categories: Sars coronavirus | Single protein | Li, J. | Rao, Z. | Shen, W. | Xue, X. | Yang, H. | Zhao, Q. | 3IH | Main protease | Michael acceptor n3 | Sars-cov
