Cargando…
Targeted Covalent Inhibitors Allosterically Deactivate the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal Sites
[Image: see text] For using targeted covalent inhibitors (TCIs) as anticancer and antiviral drugs, we establish that the model compounds PCMPS (p-chloromercuriphenyl sulfate) and PCMB (p-chloromercuribenzoate) are inhibitors of the DEDDh family of exonucleases. The underlying mechanism is analyzed b...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
|
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715546/ https://www.ncbi.nlm.nih.gov/pubmed/34977900 http://dx.doi.org/10.1021/jacsau.1c00420 |
_version_ | 1784624148098580480 |
---|---|
author | Huang, Kuan-Wei Chen, Jing-Wen Hua, Tzu-Yu Chu, Yu-Yu Chiu, Tsai-Yuan Liu, Jung-Yu Tu, Chun-I Hsu, Kai-Cheng Kao, Ya-Ting Chu, Jhih-Wei Hsiao, Yu-Yuan |
author_facet | Huang, Kuan-Wei Chen, Jing-Wen Hua, Tzu-Yu Chu, Yu-Yu Chiu, Tsai-Yuan Liu, Jung-Yu Tu, Chun-I Hsu, Kai-Cheng Kao, Ya-Ting Chu, Jhih-Wei Hsiao, Yu-Yuan |
author_sort | Huang, Kuan-Wei |
collection | PubMed |
description | [Image: see text] For using targeted covalent inhibitors (TCIs) as anticancer and antiviral drugs, we establish that the model compounds PCMPS (p-chloromercuriphenyl sulfate) and PCMB (p-chloromercuribenzoate) are inhibitors of the DEDDh family of exonucleases. The underlying mechanism is analyzed by X-ray crystallography, activity/nucleic acid-binding assays, and all-atom molecular dynamics (MD) simulations. The first TCI-complexed structures of a DEDDh enzyme, the Lassa fever virus NP exonuclease (NPexo), are resolved to elucidate that the Cys409 binding site is away from the active site and the RNA-binding lid. The NPexo C409A structures indicate Cys461 as the alternative distal site for obstructing the equally active mutant. All-atom MD simulations of the wild type and mutant NPexos in explicit solvent uncover an allosteric inhibition mechanism that the local perturbation induced by PCMPS sulfonate propagates to impact the RNA-binding lid conformation. Binding assay studies confirm that PCMPS does affect the RNA binding of NPexo. The predicted relative potency between PCMPS and PCMB is also in line with experiments. The structural data and inhibition mechanism established in this work provide an important molecular basis for the drug development of TCIs. |
format | Online Article Text |
id | pubmed-8715546 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-87155462021-12-30 Targeted Covalent Inhibitors Allosterically Deactivate the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal Sites Huang, Kuan-Wei Chen, Jing-Wen Hua, Tzu-Yu Chu, Yu-Yu Chiu, Tsai-Yuan Liu, Jung-Yu Tu, Chun-I Hsu, Kai-Cheng Kao, Ya-Ting Chu, Jhih-Wei Hsiao, Yu-Yuan JACS Au [Image: see text] For using targeted covalent inhibitors (TCIs) as anticancer and antiviral drugs, we establish that the model compounds PCMPS (p-chloromercuriphenyl sulfate) and PCMB (p-chloromercuribenzoate) are inhibitors of the DEDDh family of exonucleases. The underlying mechanism is analyzed by X-ray crystallography, activity/nucleic acid-binding assays, and all-atom molecular dynamics (MD) simulations. The first TCI-complexed structures of a DEDDh enzyme, the Lassa fever virus NP exonuclease (NPexo), are resolved to elucidate that the Cys409 binding site is away from the active site and the RNA-binding lid. The NPexo C409A structures indicate Cys461 as the alternative distal site for obstructing the equally active mutant. All-atom MD simulations of the wild type and mutant NPexos in explicit solvent uncover an allosteric inhibition mechanism that the local perturbation induced by PCMPS sulfonate propagates to impact the RNA-binding lid conformation. Binding assay studies confirm that PCMPS does affect the RNA binding of NPexo. The predicted relative potency between PCMPS and PCMB is also in line with experiments. The structural data and inhibition mechanism established in this work provide an important molecular basis for the drug development of TCIs. American Chemical Society 2021-11-16 /pmc/articles/PMC8715546/ /pubmed/34977900 http://dx.doi.org/10.1021/jacsau.1c00420 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Huang, Kuan-Wei Chen, Jing-Wen Hua, Tzu-Yu Chu, Yu-Yu Chiu, Tsai-Yuan Liu, Jung-Yu Tu, Chun-I Hsu, Kai-Cheng Kao, Ya-Ting Chu, Jhih-Wei Hsiao, Yu-Yuan Targeted Covalent Inhibitors Allosterically Deactivate the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal Sites |
title | Targeted Covalent Inhibitors Allosterically Deactivate
the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal
Sites |
title_full | Targeted Covalent Inhibitors Allosterically Deactivate
the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal
Sites |
title_fullStr | Targeted Covalent Inhibitors Allosterically Deactivate
the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal
Sites |
title_full_unstemmed | Targeted Covalent Inhibitors Allosterically Deactivate
the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal
Sites |
title_short | Targeted Covalent Inhibitors Allosterically Deactivate
the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal
Sites |
title_sort | targeted covalent inhibitors allosterically deactivate
the deddh lassa fever virus np exonuclease from alternative distal
sites |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715546/ https://www.ncbi.nlm.nih.gov/pubmed/34977900 http://dx.doi.org/10.1021/jacsau.1c00420 |
work_keys_str_mv | AT huangkuanwei targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT chenjingwen targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT huatzuyu targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT chuyuyu targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT chiutsaiyuan targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT liujungyu targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT tuchuni targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT hsukaicheng targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT kaoyating targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT chujhihwei targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites AT hsiaoyuyuan targetedcovalentinhibitorsallostericallydeactivatethededdhlassafevervirusnpexonucleasefromalternativedistalsites |