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Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus nsp5 Protease (3CLpro): IMPLICATIONS FOR nsp5 REGULATION AND THE DEVELOPMENT OF ANTIVIRALS.
J Biol Chem. 2015 Aug 07; 290(32):19403-22.JB

Abstract

All coronaviruses, including the recently emerged Middle East respiratory syndrome coronavirus (MERS-CoV) from the β-CoV subgroup, require the proteolytic activity of the nsp5 protease (also known as 3C-like protease, 3CL(pro)) during virus replication, making it a high value target for the development of anti-coronavirus therapeutics. Kinetic studies indicate that in contrast to 3CL(pro) from other β-CoV 2c members, including HKU4 and HKU5, MERS-CoV 3CL(pro) is less efficient at processing a peptide substrate due to MERS-CoV 3CL(pro) being a weakly associated dimer. Conversely, HKU4, HKU5, and SARS-CoV 3CL(pro) enzymes are tightly associated dimers. Analytical ultracentrifugation studies support that MERS-CoV 3CL(pro) is a weakly associated dimer (Kd ∼52 μm) with a slow off-rate. Peptidomimetic inhibitors of MERS-CoV 3CL(pro) were synthesized and utilized in analytical ultracentrifugation experiments and demonstrate that MERS-CoV 3CL(pro) undergoes significant ligand-induced dimerization. Kinetic studies also revealed that designed reversible inhibitors act as activators at a low compound concentration as a result of induced dimerization. Primary sequence comparisons and x-ray structural analyses of two MERS-CoV 3CLpro and inhibitor complexes, determined to 1.6 Å, reveal remarkable structural similarity of the dimer interface with 3CL(pro) from HKU4-CoV and HKU5-CoV. Despite this structural similarity, substantial differences in the dimerization ability suggest that long range interactions by the nonconserved amino acids distant from the dimer interface may control MERS-CoV 3CL(pro) dimerization. Activation of MERS-CoV 3CL(pro) through ligand-induced dimerization appears to be unique within the genogroup 2c and may potentially increase the complexity in the development of MERS-CoV 3CL(pro) inhibitors as antiviral agents.

Authors+Show Affiliations

From the Departments of Biological Sciences and.Chemistry, Purdue University, West Lafayette, Indiana 47907.From the Departments of Biological Sciences and.Chemistry, Purdue University, West Lafayette, Indiana 47907.Chemistry, Purdue University, West Lafayette, Indiana 47907.the Bindley Bioscience Center, Purdue University, West Lafayette, Indiana 47907, and.Chemistry, Purdue University, West Lafayette, Indiana 47907.the Departments of Pediatrics and Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.From the Departments of Biological Sciences and Chemistry, Purdue University, West Lafayette, Indiana 47907, amesecar@purdue.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

26055715

Citation

Tomar, Sakshi, et al. "Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus Nsp5 Protease (3CLpro): IMPLICATIONS for Nsp5 REGULATION and the DEVELOPMENT of ANTIVIRALS." The Journal of Biological Chemistry, vol. 290, no. 32, 2015, pp. 19403-22.
Tomar S, Johnston ML, St John SE, et al. Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus nsp5 Protease (3CLpro): IMPLICATIONS FOR nsp5 REGULATION AND THE DEVELOPMENT OF ANTIVIRALS. J Biol Chem. 2015;290(32):19403-22.
Tomar, S., Johnston, M. L., St John, S. E., Osswald, H. L., Nyalapatla, P. R., Paul, L. N., Ghosh, A. K., Denison, M. R., & Mesecar, A. D. (2015). Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus nsp5 Protease (3CLpro): IMPLICATIONS FOR nsp5 REGULATION AND THE DEVELOPMENT OF ANTIVIRALS. The Journal of Biological Chemistry, 290(32), 19403-22. https://doi.org/10.1074/jbc.M115.651463
Tomar S, et al. Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus Nsp5 Protease (3CLpro): IMPLICATIONS for Nsp5 REGULATION and the DEVELOPMENT of ANTIVIRALS. J Biol Chem. 2015 Aug 7;290(32):19403-22. PubMed PMID: 26055715.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ligand-induced Dimerization of Middle East Respiratory Syndrome (MERS) Coronavirus nsp5 Protease (3CLpro): IMPLICATIONS FOR nsp5 REGULATION AND THE DEVELOPMENT OF ANTIVIRALS. AU - Tomar,Sakshi, AU - Johnston,Melanie L, AU - St John,Sarah E, AU - Osswald,Heather L, AU - Nyalapatla,Prasanth R, AU - Paul,Lake N, AU - Ghosh,Arun K, AU - Denison,Mark R, AU - Mesecar,Andrew D, Y1 - 2015/06/08/ PY - 2015/03/11/received PY - 2015/6/10/entrez PY - 2015/6/10/pubmed PY - 2015/11/6/medline KW - MERS-CoV 3CLpro KW - X-ray crystallography KW - analytical ultracentrifugation KW - enzyme inactivation KW - enzyme inhibitor KW - enzyme kinetics KW - ligand-induced dimerization KW - monomer-dimer equilibrium KW - viral protease KW - β-CoV SP - 19403 EP - 22 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 290 IS - 32 N2 - All coronaviruses, including the recently emerged Middle East respiratory syndrome coronavirus (MERS-CoV) from the β-CoV subgroup, require the proteolytic activity of the nsp5 protease (also known as 3C-like protease, 3CL(pro)) during virus replication, making it a high value target for the development of anti-coronavirus therapeutics. Kinetic studies indicate that in contrast to 3CL(pro) from other β-CoV 2c members, including HKU4 and HKU5, MERS-CoV 3CL(pro) is less efficient at processing a peptide substrate due to MERS-CoV 3CL(pro) being a weakly associated dimer. Conversely, HKU4, HKU5, and SARS-CoV 3CL(pro) enzymes are tightly associated dimers. Analytical ultracentrifugation studies support that MERS-CoV 3CL(pro) is a weakly associated dimer (Kd ∼52 μm) with a slow off-rate. Peptidomimetic inhibitors of MERS-CoV 3CL(pro) were synthesized and utilized in analytical ultracentrifugation experiments and demonstrate that MERS-CoV 3CL(pro) undergoes significant ligand-induced dimerization. Kinetic studies also revealed that designed reversible inhibitors act as activators at a low compound concentration as a result of induced dimerization. Primary sequence comparisons and x-ray structural analyses of two MERS-CoV 3CLpro and inhibitor complexes, determined to 1.6 Å, reveal remarkable structural similarity of the dimer interface with 3CL(pro) from HKU4-CoV and HKU5-CoV. Despite this structural similarity, substantial differences in the dimerization ability suggest that long range interactions by the nonconserved amino acids distant from the dimer interface may control MERS-CoV 3CL(pro) dimerization. Activation of MERS-CoV 3CL(pro) through ligand-induced dimerization appears to be unique within the genogroup 2c and may potentially increase the complexity in the development of MERS-CoV 3CL(pro) inhibitors as antiviral agents. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/26055715/Ligand_induced_Dimerization_of_Middle_East_Respiratory_Syndrome__MERS__Coronavirus_nsp5_Protease__3CLpro_:_IMPLICATIONS_FOR_nsp5_REGULATION_AND_THE_DEVELOPMENT_OF_ANTIVIRALS_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=26055715 DB - PRIME DP - Unbound Medicine ER -