Abstract
Papain-like protease (PLpro) from SARS-CoV-2 plays essential roles in the replication cycle of the virus. In particular, it preferentially interacts with and cleaves human interferon-stimulated gene 15 (hISG15) to suppress the innate immune response of the host. We used small-angle X-ray and neutron scattering combined with computational techniques to study the mechanism of interaction of SARS-CoV-2 PLpro with hISG15. We showed that hISG15 undergoes a transition from an extended to a compact state after binding to PLpro, a conformation that has not been previously observed in complexes of SARS-CoV-2 PLpro with ISG15 from other species. Furthermore, computational analysis showed significant conformational flexibility in the ISG15 N-terminal domain, suggesting that it is weakly bound to PLpro and supports a binding mechanism that is dominated by the C-terminal ISG15 domain. This study fundamentally improves our understanding of the SARS-CoV-2 deISGylation complex that will help guide development of COVID-19 therapeutics targeting this complex.
| Original language | English |
|---|---|
| Pages (from-to) | 5608-5615 |
| Number of pages | 8 |
| Journal | Journal of Physical Chemistry Letters |
| Volume | 12 |
| Issue number | 23 |
| DOIs | |
| State | Published - Jun 17 2021 |
Funding
This research was supported by the U.S. Department of Energy (DOE) Office of Science through the National Virtual Biotechnology Laboratory (NVBL), a consortium of DOE National Laboratories focused on response to COVID-19, with funding provided by the Coronavirus CARES Act. Preliminary protein expression and purification studies were supported by the Laboratory Directed Research and Development Program at Oak Ridge National Laboratory (ORNL). The SARS-CoV-2 PLpro C111S expression plasmid (pMCSG53) was provided by Dr. Andrzej Joachimiak (Argonne National Laboratory, Lemont, IL) with support from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Contract HHSN272201700060C. Bio-SANS is supported by ORNL’s Center for Structural Molecular Biology funded by the DOE Office of Biological and Environmental Research. This research used resources at the High Flux Isotope Reactor and Spallation Neutron Source, a DOE Office of Science User Facility operated by the Oak Ridge National Laboratory. SAXS was conducted at the Advanced Light Source (ALS), a national user facility operated by Lawrence Berkeley National Laboratory on behalf of the DOE, Office of Basic Energy Sciences, through the Integrated Diffraction Analysis Technologies (IDAT) program, supported by DOE Office of Biological and Environmental Research. Additional support comes from National Institutes of Health Project ALS-ENABLE (P30 GM124169) and High-End Instrumentation Grant S10OD018483. L.C. acknowledges support from the Second Target Station, a DOE Office of Science User Facilities Construction Project at Oak Ridge National Laboratory. This manuscript has been authored by UT-Battelle, LLC, under Contract DE-AC05-00OR22725 with the DOE. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this manuscript, or allow others to do so, for U.S. Government purposes. The DOE will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan ( http://energy.gov/downloads/doe-public-access-plan ).
| Funders | Funder number |
|---|---|
| DOE National Laboratories | |
| National Virtual Biotechnology Laboratory | |
| Second Target Station | |
| National Institutes of Health | |
| U.S. Department of Energy | |
| U.S. Department of Health and Human Services | HHSN272201700060C |
| NIH Office of the Director | S10OD018483 |
| National Institute of General Medical Sciences | P30GM124169 |
| National Institute of Allergy and Infectious Diseases | |
| Office of Science | DE-AC05-00OR22725 |
| Basic Energy Sciences | P30 GM124169 |
| Biological and Environmental Research | |
| Argonne National Laboratory | |
| Oak Ridge National Laboratory |
