We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Dual domain recognition determines SARS-CoV-2 PLpro selectivity for human ISG15 and K48-linked di-ubiquitin.
- Authors
Wydorski, Pawel M.; Osipiuk, Jerzy; Lanham, Benjamin T.; Tesar, Christine; Endres, Michael; Engle, Elizabeth; Jedrzejczak, Robert; Mullapudi, Vishruth; Michalska, Karolina; Fidelis, Krzysztof; Fushman, David; Joachimiak, Andrzej; Joachimiak, Lukasz A.
- Abstract
The Papain-like protease (PLpro) is a domain of a multi-functional, non-structural protein 3 of coronaviruses. PLpro cleaves viral polyproteins and posttranslational conjugates with poly-ubiquitin and protective ISG15, composed of two ubiquitin-like (UBL) domains. Across coronaviruses, PLpro showed divergent selectivity for recognition and cleavage of posttranslational conjugates despite sequence conservation. We show that SARS-CoV-2 PLpro binds human ISG15 and K48-linked di-ubiquitin (K48-Ub2) with nanomolar affinity and detect alternate weaker-binding modes. Crystal structures of untethered PLpro complexes with ISG15 and K48-Ub2 combined with solution NMR and cross-linking mass spectrometry revealed how the two domains of ISG15 or K48-Ub2 are differently utilized in interactions with PLpro. Analysis of protein interface energetics predicted differential binding stabilities of the two UBL/Ub domains that were validated experimentally. We emphasize how substrate recognition can be tuned to cleave specifically ISG15 or K48-Ub2 modifications while retaining capacity to cleave mono-Ub conjugates. These results highlight alternative druggable surfaces that would inhibit PLpro function. Understanding mechanisms of PLpro substrate selectivity offers new ways to decouple substrate activities and will inform new therapeutic strategies. Here, the authors use multi-disciplinary approaches to uncover how PLpro from SARS-CoV-2 can discriminate between different substrates.
- Subjects
SARS-CoV-2; MASS spectrometry; PROTEIN analysis; CORONAVIRUSES; CRYSTAL structure; ADP-ribosylation
- Publication
Nature Communications, 2023, Vol 14, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-023-38031-5