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Product release is rate-limiting for catalytic processing by the Dengue virus protease
Dengue Virus (DENV) is the most prevalent global arbovirus, yet despite an increasing burden to health care there are currently no therapeutics available to treat infection. A potential target for antiviral drugs is the two-component viral protease NS2B-NS3pro, which is essential for viral replicati...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126634/ https://www.ncbi.nlm.nih.gov/pubmed/27897196 http://dx.doi.org/10.1038/srep37539 |
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author | Shannon, A. E. Pedroso, M. M. Chappell, K. J. Watterson, D. Liebscher, S. Kok, W. M. Fairlie, D. P. Schenk, G. Young, P. R. |
author_facet | Shannon, A. E. Pedroso, M. M. Chappell, K. J. Watterson, D. Liebscher, S. Kok, W. M. Fairlie, D. P. Schenk, G. Young, P. R. |
author_sort | Shannon, A. E. |
collection | PubMed |
description | Dengue Virus (DENV) is the most prevalent global arbovirus, yet despite an increasing burden to health care there are currently no therapeutics available to treat infection. A potential target for antiviral drugs is the two-component viral protease NS2B-NS3pro, which is essential for viral replication. Interactions between the two components have been investigated here by probing the effect on the rate of enzyme catalysis of key mutations in a mobile loop within NS2B that is located at the interface of the two components. Steady-state kinetic assays indicated that the mutations greatly affect catalytic turnover. However, single turnover and fluorescence experiments have revealed that the mutations predominantly affect product release rather than substrate binding. Fluorescence analysis also indicated that the addition of substrate triggers a near-irreversible change in the enzyme conformation that activates the catalytic centre. Based on this mechanistic insight, we propose that residues within the mobile loop of NS2B control product release and present a new target for design of potent Dengue NS2B-NS3 protease inhibitors. |
format | Online Article Text |
id | pubmed-5126634 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-51266342016-12-09 Product release is rate-limiting for catalytic processing by the Dengue virus protease Shannon, A. E. Pedroso, M. M. Chappell, K. J. Watterson, D. Liebscher, S. Kok, W. M. Fairlie, D. P. Schenk, G. Young, P. R. Sci Rep Article Dengue Virus (DENV) is the most prevalent global arbovirus, yet despite an increasing burden to health care there are currently no therapeutics available to treat infection. A potential target for antiviral drugs is the two-component viral protease NS2B-NS3pro, which is essential for viral replication. Interactions between the two components have been investigated here by probing the effect on the rate of enzyme catalysis of key mutations in a mobile loop within NS2B that is located at the interface of the two components. Steady-state kinetic assays indicated that the mutations greatly affect catalytic turnover. However, single turnover and fluorescence experiments have revealed that the mutations predominantly affect product release rather than substrate binding. Fluorescence analysis also indicated that the addition of substrate triggers a near-irreversible change in the enzyme conformation that activates the catalytic centre. Based on this mechanistic insight, we propose that residues within the mobile loop of NS2B control product release and present a new target for design of potent Dengue NS2B-NS3 protease inhibitors. Nature Publishing Group 2016-11-29 /pmc/articles/PMC5126634/ /pubmed/27897196 http://dx.doi.org/10.1038/srep37539 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Shannon, A. E. Pedroso, M. M. Chappell, K. J. Watterson, D. Liebscher, S. Kok, W. M. Fairlie, D. P. Schenk, G. Young, P. R. Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title | Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title_full | Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title_fullStr | Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title_full_unstemmed | Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title_short | Product release is rate-limiting for catalytic processing by the Dengue virus protease |
title_sort | product release is rate-limiting for catalytic processing by the dengue virus protease |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126634/ https://www.ncbi.nlm.nih.gov/pubmed/27897196 http://dx.doi.org/10.1038/srep37539 |
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