Long-Range Cooperative Interactions Modulate Dimerization in SARS 3CLpro
- 18 November 2006
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 45 (50), 14908-14916
- https://doi.org/10.1021/bi0616302
Abstract
Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CLpro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CLpro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry43, 4906−4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 Å away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.Keywords
This publication has 34 references indexed in Scilit:
- The N-terminal octapeptide acts as a dimerization inhibitor of SARS coronavirus 3C-like proteinaseBiochemical and Biophysical Research Communications, 2006
- Biosynthesis, Purification, and Substrate Specificity of Severe Acute Respiratory Syndrome Coronavirus 3C-like ProteinasePublished by Elsevier BV ,2004
- The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitorProceedings of the National Academy of Sciences of the United States of America, 2003
- On the analysis of protein self-association by sedimentation velocity analytical ultracentrifugationAnalytical Biochemistry, 2003
- Coronavirus Main Proteinase (3CL pro ) Structure: Basis for Design of Anti-SARS DrugsScience, 2003
- A Novel Coronavirus Associated with Severe Acute Respiratory SyndromeNew England Journal of Medicine, 2003
- Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia: a prospective studyThe Lancet, 2003
- Coronavirus as a possible cause of severe acute respiratory syndromeThe Lancet, 2003
- Structure of coronavirus main proteinase reveals combination of a chymotrypsin fold with an extra alpha-helical domainThe EMBO Journal, 2002
- Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation ModelingBiophysical Journal, 2000