Self-association and conformational variation of NS5A domain 1 of hepatitis C virus
Direct-acting antivirals (DAAs) targeting the non-structural 5A (NS5A) protein of the hepatitis C virus (HCV) are crucial drugs that have shown exceptional clinical success in patients. However, their mode of action (MoA) remains unclear, and drug-resistant HCV strains are rapidly emerging. It is cr...
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sg-ntu-dr.10356-1425892020-06-25T02:31:12Z Self-association and conformational variation of NS5A domain 1 of hepatitis C virus Beldar, Serap Manimekalai, Malathy Sony Subramanian Cho, Nam-Joon Baek, Kwanghee Grüber, Gerhard Yoon, Ho Sup School of Materials Science and Engineering School of Biological Sciences Science::Biological sciences Hepatitis C Virus NS5A Protein Direct-acting antivirals (DAAs) targeting the non-structural 5A (NS5A) protein of the hepatitis C virus (HCV) are crucial drugs that have shown exceptional clinical success in patients. However, their mode of action (MoA) remains unclear, and drug-resistant HCV strains are rapidly emerging. It is critical to characterize the behaviour of the NS5A protein in solution, which can facilitate the development of new classes of inhibitors or improve the efficacy of the currently available DAAs. Using biophysical methods, including dynamic light scattering, size exclusion chromatography and chemical cross-linking experiments, we showed that the NS5A domain 1 from genotypes 1b and 1a of the HCV intrinsically self-associated and existed as a heterogeneous mixture in solution. Interestingly, the NS5A domain 1 from genotypes 1b and 1a exhibited different dynamic equilibria of monomers to higher-order structures. Using small-angle X-ray scattering, we studied the structural dynamics of the various states of the NS5A domain 1 in solution. We also tested the effect of daclatasvir (DCV), the most prominent DAA, on self-association of the wild and DCV-resistant mutant (Y93H) NS5A domain 1 proteins, and demonstrated that DCV induced the formation of large and irreversible protein aggregates that eventually precipitated out. This study highlights the conformational variability of the NS5A domain 1 of HCV, which may be an intrinsic structural behaviour of the HCV NS5A domain 1 in solution. MOE (Min. of Education, S’pore) 2020-06-25T02:31:11Z 2020-06-25T02:31:11Z 2018 Journal Article Beldar, S., Manimekalai, M. S. S., Cho, N.-J., Baek, K., Grüber, G., & Yoon, H. S. (2018). Self-association and conformational variation of NS5A domain 1 of hepatitis C virus. Journal of general virology, 99(2), 194–208. doi:10.1099/jgv.0.001000 0022-1317 https://hdl.handle.net/10356/142589 10.1099/jgv.0.001000 29300159 2-s2.0-85041809007 2 99 194 208 en Journal of general virology © 2018 The Authors. All rights reserved. |
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Science::Biological sciences Hepatitis C Virus NS5A Protein Beldar, Serap Manimekalai, Malathy Sony Subramanian Cho, Nam-Joon Baek, Kwanghee Grüber, Gerhard Yoon, Ho Sup Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
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Direct-acting antivirals (DAAs) targeting the non-structural 5A (NS5A) protein of the hepatitis C virus (HCV) are crucial drugs that have shown exceptional clinical success in patients. However, their mode of action (MoA) remains unclear, and drug-resistant HCV strains are rapidly emerging. It is critical to characterize the behaviour of the NS5A protein in solution, which can facilitate the development of new classes of inhibitors or improve the efficacy of the currently available DAAs. Using biophysical methods, including dynamic light scattering, size exclusion chromatography and chemical cross-linking experiments, we showed that the NS5A domain 1 from genotypes 1b and 1a of the HCV intrinsically self-associated and existed as a heterogeneous mixture in solution. Interestingly, the NS5A domain 1 from genotypes 1b and 1a exhibited different dynamic equilibria of monomers to higher-order structures. Using small-angle X-ray scattering, we studied the structural dynamics of the various states of the NS5A domain 1 in solution. We also tested the effect of daclatasvir (DCV), the most prominent DAA, on self-association of the wild and DCV-resistant mutant (Y93H) NS5A domain 1 proteins, and demonstrated that DCV induced the formation of large and irreversible protein aggregates that eventually precipitated out. This study highlights the conformational variability of the NS5A domain 1 of HCV, which may be an intrinsic structural behaviour of the HCV NS5A domain 1 in solution. |
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School of Materials Science and Engineering |
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School of Materials Science and Engineering Beldar, Serap Manimekalai, Malathy Sony Subramanian Cho, Nam-Joon Baek, Kwanghee Grüber, Gerhard Yoon, Ho Sup |
format |
Article |
author |
Beldar, Serap Manimekalai, Malathy Sony Subramanian Cho, Nam-Joon Baek, Kwanghee Grüber, Gerhard Yoon, Ho Sup |
author_sort |
Beldar, Serap |
title |
Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
title_short |
Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
title_full |
Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
title_fullStr |
Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
title_full_unstemmed |
Self-association and conformational variation of NS5A domain 1 of hepatitis C virus |
title_sort |
self-association and conformational variation of ns5a domain 1 of hepatitis c virus |
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2020 |
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https://hdl.handle.net/10356/142589 |
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1681057911712776192 |