tetano
Editor, Senior Moderator
Nat Commun
. 2023 Apr 25;14(1):2366.
doi: 10.1038/s41467-023-38031-5. Dual domain recognition determines SARS-CoV-2 PLpro selectivity for human ISG15 and K48-linked di-ubiquitin
Pawel M Wydorski[SUP] #[/SUP][SUP] 1 2 [/SUP], Jerzy Osipiuk[SUP] #[/SUP][SUP] 3 4 [/SUP], Benjamin T Lanham[SUP] #[/SUP][SUP] 5 [/SUP], Christine Tesar[SUP] 3 4 [/SUP], Michael Endres[SUP] 3 4 [/SUP], Elizabeth Engle[SUP] 5 [/SUP], Robert Jedrzejczak[SUP] 3 4 [/SUP], Vishruth Mullapudi[SUP] 2 [/SUP], Karolina Michalska[SUP] 3 4 [/SUP], Krzysztof Fidelis[SUP] 6 [/SUP], David Fushman[SUP] 7 [/SUP], Andrzej Joachimiak[SUP] 8 9 10 [/SUP], Lukasz A Joachimiak[SUP] 11 12 [/SUP]
Affiliations
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-Ub[SUB]2[/SUB]) with nanomolar affinity and detect alternate weaker-binding modes. Crystal structures of untethered PLpro complexes with ISG15 and K48-Ub[SUB]2[/SUB] combined with solution NMR and cross-linking mass spectrometry revealed how the two domains of ISG15 or K48-Ub[SUB]2[/SUB] 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-Ub[SUB]2[/SUB] modifications while retaining capacity to cleave mono-Ub conjugates. These results highlight alternative druggable surfaces that would inhibit PLpro function.
. 2023 Apr 25;14(1):2366.
doi: 10.1038/s41467-023-38031-5. Dual domain recognition determines SARS-CoV-2 PLpro selectivity for human ISG15 and K48-linked di-ubiquitin
Pawel M Wydorski[SUP] #[/SUP][SUP] 1 2 [/SUP], Jerzy Osipiuk[SUP] #[/SUP][SUP] 3 4 [/SUP], Benjamin T Lanham[SUP] #[/SUP][SUP] 5 [/SUP], Christine Tesar[SUP] 3 4 [/SUP], Michael Endres[SUP] 3 4 [/SUP], Elizabeth Engle[SUP] 5 [/SUP], Robert Jedrzejczak[SUP] 3 4 [/SUP], Vishruth Mullapudi[SUP] 2 [/SUP], Karolina Michalska[SUP] 3 4 [/SUP], Krzysztof Fidelis[SUP] 6 [/SUP], David Fushman[SUP] 7 [/SUP], Andrzej Joachimiak[SUP] 8 9 10 [/SUP], Lukasz A Joachimiak[SUP] 11 12 [/SUP]
Affiliations
- PMID: 37185902
- DOI: 10.1038/s41467-023-38031-5
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-Ub[SUB]2[/SUB]) with nanomolar affinity and detect alternate weaker-binding modes. Crystal structures of untethered PLpro complexes with ISG15 and K48-Ub[SUB]2[/SUB] combined with solution NMR and cross-linking mass spectrometry revealed how the two domains of ISG15 or K48-Ub[SUB]2[/SUB] 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-Ub[SUB]2[/SUB] modifications while retaining capacity to cleave mono-Ub conjugates. These results highlight alternative druggable surfaces that would inhibit PLpro function.