Activity, Specificity, and Probe Design for the Smallpox Virus Protease K7L

Aleshin, Alexander E.; Drąg, Marcin; Gombosuren, Naran; Ge, Wei; Mikolajczyk, Jowita; Satterthwait, Arnold C.; Strongin, Alex Y.; Liddington, Robert; Salvesen, Guy S.

doi:10.1074/jbc.m112.388678

Cited by 17 publications

(28 citation statements)

References 46 publications

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“…Another possible application of our inhibitors is their use as selective ABPs to visualize SARS-CoV-2 PLpro activity in cells or even in COVID-19 diagnostics. Similar studies have already been conducted toward the use of ABPs for smallpox K7L protease, ZIKA, WNW, or dengue viruses proteases (34)(35)(36).…”

Section: Fig 5 a Molecular Basis For The Observed Ub/ubl Processingmentioning

confidence: 71%

Activity profiling and crystal structures of inhibitor-bound SARS-CoV-2 papain-like protease: A framework for anti–COVID-19 drug design

et al. 2020

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Viral papain-like cysteine protease (PLpro, NSP3) is essential for SARS-CoV-2 replication and represents a promising target for the development of antiviral drugs. Here, we used a combinatorial substrate library and performed comprehensive activity profiling of SARS-CoV-2 PLpro. On the scaffold of the best hits from positional scanning, we designed optimal fluorogenic substrates and irreversible inhibitors with a high degree of selectivity for SARS PLpro. We determined crystal structures of two of these inhibitors in complex with SARS-CoV-2 PLpro that reveals their inhibitory mechanisms and provides a molecular basis for the observed substrate specificity profiles. Last, we demonstrate that SARS-CoV-2 PLpro harbors deISGylating activity similar to SARSCoV-1 PLpro but its ability to hydrolyze K48-linked Ub chains is diminished, which our sequence and structure analysis provides a basis for. Together, this work has revealed the molecular rules governing PLpro substrate specificity and provides a framework for development of inhibitors with potential therapeutic value or drug repurposing.

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Section: Fig 5 a Molecular Basis For The Observed Ub/ubl Processingmentioning

confidence: 71%

Activity profiling and crystal structures of inhibitor-bound SARS-CoV-2 papain-like protease: A framework for anti–COVID-19 drug design

et al. 2020

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“…We propose that a conformational change in flMCP CTD improves cleavage site accessibility upon assembly, but we cannot rule out different scenarios. Regulation of virion processing has also been described for other DJR viruses encoding clan CE cysteine proteases, such as smallpox ( 34 ) and adenovirus ( 35 ). Whether MVP can be stimulated by cofactors or is affected by oligomerization remains open.…”

Section: Discussionmentioning

confidence: 92%

Capsid protein structure, self-assembly, and processing reveal morphogenesis of the marine virophage mavirus

Born

Reuter

Mersdorf

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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SignificanceVirophages are parasites of giant viruses within protists. They reduce giant virus production and increase host cell survival. They provide a defense system for protists against giant viruses in diverse environments, likely with ecological relevance for protist populations. To understand the remarkable virophage life cycle, it is crucial to investigate how they assemble into infectious particles and which processes require interactions with giant virus and host. We examined the marine virophage mavirus to show that its major and minor capsid proteins assemble into virus-like particles in the absence of specific host or viral factors. Subsequently, the virophage-encoded protease processes the major capsid protein to prepare virions for infection.

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“… 42 , 43 Similarly, VAR F1L may be a valid target for development of antiviral therapeutics. Current strategies for the development of antiviral therapeutics against VAR have relied mostly on the use of homologous target molecules from VV, monkeypox or ectromelia virus, with only a small number of studies utilizing VAR-encoded proteins, 44 , 45 , 46 and even fewer studies using their structures. 47 , 48 The marked differences between VAR and VV F1L suggest that studies of VAR proteins may provide unexpected avenues for therapeutic intervention that may not be predicted based on available data from the closely related and much better understood VV.…”

Section: Discussionmentioning

confidence: 99%

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

et al. 2015

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Subversion of host cell apoptosis is an important survival strategy for viruses to ensure their own proliferation and survival. Certain viruses express proteins homologous in sequence, structure and function to mammalian pro-survival B-cell lymphoma 2 (Bcl-2) proteins, which prevent rapid clearance of infected host cells. In vaccinia virus (VV), the virulence factor F1L was shown to be a potent inhibitor of apoptosis that functions primarily be engaging pro-apoptotic Bim. Variola virus (VAR), the causative agent of smallpox, harbors a homolog of F1L of unknown function. We show that VAR F1L is a potent inhibitor of apoptosis, and unlike all other characterized anti-apoptotic Bcl-2 family members lacks affinity for the Bim Bcl-2 homology 3 (BH3) domain. Instead, VAR F1L engages Bid BH3 as well as Bak and Bax BH3 domains. Unlike its VV homolog, variola F1L only protects against Bax-mediated apoptosis in cellular assays. Crystal structures of variola F1L bound to Bid and Bak BH3 domains reveal that variola F1L forms a domain-swapped Bcl-2 fold, which accommodates Bid and Bak BH3 in the canonical Bcl-2-binding groove, in a manner similar to VV F1L. Despite the observed conservation of structure and sequence, variola F1L inhibits apoptosis using a startlingly different mechanism compared with its VV counterpart. Our results suggest that unlike during VV infection, Bim neutralization may not be required during VAR infection. As molecular determinants for the human-specific tropism of VAR remain essentially unknown, identification of a different mechanism of action and utilization of host factors used by a VAR virulence factor compared with its VV homolog suggest that studying VAR directly may be essential to understand its unique tropism.

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Activity, Specificity, and Probe Design for the Smallpox Virus Protease K7L

Cited by 17 publications

References 46 publications

Activity profiling and crystal structures of inhibitor-bound SARS-CoV-2 papain-like protease: A framework for anti–COVID-19 drug design

Activity profiling and crystal structures of inhibitor-bound SARS-CoV-2 papain-like protease: A framework for anti–COVID-19 drug design

Capsid protein structure, self-assembly, and processing reveal morphogenesis of the marine virophage mavirus

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

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