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Published Online May 13, 2003
Science DOI: 10.1126/science.1085658

Reports

Submitted on April 14, 2003
Accepted on May 12, 2003

Coronavirus Main Proteinase (3CLpro) Structure: Basis for Design of Anti-SARS Drugs

Kanchan Anand 1, John Ziebuhr 2, Parvesh Wadhwani 3, Jeroen R. Mesters 1, Rolf Hilgenfeld 4*

1 Institute of Biochemistry, University of Lübeck, D-23538 Lübeck, Germany; Institute of Molecular Biotechnology, D-07745 Jena, Germany.
2 Institute of Virology and Immunology, University of Würzburg, D-97078 Würzburg, Germany.
3 Institute of Molecular Biology, University of Jena, D-07745 Jena, Germany.
4 Institute of Biochemistry, University of Lübeck, D-23538 Lübeck, Germany; Institute of Molecular Biotechnology, D-07745 Jena, Germany; Address for correspondence: Institute of Biochemistry, University of Lübeck, Ratzeburger Allee 160, D-23538 Lübeck, Germany.

* To whom correspondence should be addressed. E-mail: hilgenfeld{at}biochem.uni-luebeck.de.

A novel coronavirus has been identified as the causative agent of severe acute respiratory syndrome (SARS). The viral main proteinase (Mpro, also called 3CLpro), controlling the activities of the coronavirus replication complex, represents an attractive target for therapy. We determined crystal structures for human coronavirus (strain 229E) Mpro and for an inhibitor complex of porcine coronavirus (transmissible gastroenteritis virus, TGEV) Mpro, and constructed a homology model for SARS coronavirus (SARS-CoV) Mpro. The structures reveal a remarkable degree of conservation of the substrate-binding sites, which is further supported by recombinant SARS-CoV Mpro-mediated cleavage of a TGEV Mpro substrate. Molecular modeling suggests that available rhinovirus 3Cpro inhibitors may be modified to make them useful for SARS therapy.


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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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