2HOB

Crystal structure of SARS-CoV main protease with authentic N and C-termini in complex with a Michael acceptor N3

Summary for 2HOB

• Entry DOI: 10.2210/pdb2hob/pdb
• Related PRD ID: PRD_002214
• Descriptor: Replicase polyprotein 1ab, N-[(5-METHYLISOXAZOL-3-YL)CARBONYL]ALANYL-L-VALYL-N~1~-((1R,2Z)-4-(BENZYLOXY)-4-OXO-1-{[(3R)-2-OXOPYRROLIDIN-3-YL]METHYL}BUT-2-ENYL)-L-LEUCINAMIDE (3 entities in total)
• Functional Keywords: sars-cov, main protease, michael acceptor n3, viral protein
• Biological source: SARS coronavirus; More
• Total number of polymer chains: 2
• Total formula weight: 34557.43

Authors

Xue, X.,Yang, H.,Shen, W.,Zhao, Q.,Li, J.,Rao, Z. (deposition date: 2006-07-14, release date: 2007-04-03, Last modification date: 2024-11-06)

Primary citation

Xue, X.,Yang, H.,Shen, W.,Zhao, Q.,Li, J.,Yang, K.,Chen, C.,Jin, Y.,Bartlam, M.,Rao, Z.
Production of authentic SARS-CoV M(pro) with enhanced activity: application as a novel tag-cleavage endopeptidase for protein overproduction
J.Mol.Biol., 366:965-975, 2007

PubMed Abstract: 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.

PubMed: 17189639
DOI: 10.1016/j.jmb.2006.11.073

Experimental method

X-RAY DIFFRACTION (1.95 Å)