Structure of the NS2B-NS3 protease from Zika virus after self-cleavage

Phoo, Wint Wint; Li, Yan; Zhang, Zhenzhen; Lee, Michelle Yueqi; Loh, Ying Ru; Tan, Yen Nee; Ng, Elizabeth Yihui; Lescar, Julien; Kang, CongBao; Luo, Dahai

doi:10.1038/ncomms13410

Cited by 194 publications

(255 citation statements)

References 46 publications

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“…Not surprisingly, the fold of flaviviral NS3 proteases is also similar. The recently solved structures of ZIKV NS2B-NS3pro support this conclusion (Lei et al, 2016; Phoo et al, 2016). Moreover, the structural similarities are consistent with the potent activity of aprotinin as an active site inhibitor of both WNV and ZIKV proteases.…”

Section: Discussionmentioning

confidence: 75%

Characterization of the Zika virus two-component NS2B-NS3 protease and structure-assisted identification of allosteric small-molecule antagonists

et al. 2017

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The recent re-emergence of Zika virus (ZIKV)1, a member of the Flaviviridae family, has become a global emergency. Currently, there are no effective methods of preventing or treating ZIKV infection, which causes severe neuroimmunopathology and is particularly harmful to the developing fetuses of infected pregnant women. However, the pathology induced by ZIKV is unique among flaviviruses, and knowledge of the biology of other family members cannot easily be extrapolated to ZIKV. Thus, structure-function studies of ZIKV proteins are urgently needed to facilitate the development of effective preventative and therapeutic agents. Like other flaviviruses, ZIKV expresses an NS2B-NS3 protease, which consists of the NS2B cofactor and the NS3 protease domain and is essential for cleavage of the ZIKV polyprotein precursor and generation of fully functional viral proteins. Here, we report the enzymatic characterization of ZIKV protease, and we identify structural scaffolds for allosteric small-molecule inhibitors of this protease. Molecular modeling of the protease-inhibitor complexes suggests that these compounds bind to the druggable cavity in the NS2B-NS3 protease interface and affect productive interactions of the protease domain with its cofactor. The most potent compound demonstrated efficient inhibition of ZIKV propagation in vitro in human fetal neural progenitor cells and in vivo in SJL mice. The inhibitory scaffolds could be further developed into valuable research reagents and, ultimately, provide a roadmap for the selection of efficient inhibitors of ZIKV infection.

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Section: Discussionmentioning

confidence: 75%

Characterization of the Zika virus two-component NS2B-NS3 protease and structure-assisted identification of allosteric small-molecule antagonists

et al. 2017

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“…Construct‐dependent cleavage patterns of ZIKV NS2B CF ‐NS3 pro . (A) In a previous structural study, a linked protease with a shortened native sequence was used instead of the commonly used glycine linker (path I) . Here, the protease underwent autocatalytic cleavage at residue R101 NS2B (red letters, sequence alignment) in the NS2B C‐terminus (blue).…”

Section: Discussionmentioning

confidence: 99%

Cis autocatalytic cleavage of glycine‐linked Zika virus NS2B‐NS3 protease constructs

et al. 2019

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The flaviviral heterodimeric serine protease NS2B‐NS3, consisting of the NS3 protease domain and the NS2B co‐factor, is essential for ZIKA virus maturation and replication in cells. For in vitro studies a ‘linked’ construct, where a polyglycine linker connects NS2BCF and NS3pro, is often used. This construct undergoes autocatalytic cleavage. Here, we show that linked ZIKV NS2BCF‐NS3pro is cleaved in cis in the NS2BCF exclusively at position R95 and not at the previously proposed alternate cleavage site at residue R29 in the NS3pro. Cleavage neither affects protease stability nor activity, despite some observed differences in spectroscopic behavior. This minimally modified construct may thus be useful for future structural and functional studies of the flaviviral protease, for example when testing new inhibitors.

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“…The NS2B-NS3 protease is a high-priority drug target [17][18][19][20]. Most attempts to develop flavivirus protease inhibitors have focused on the NS3 active site with limited success, possibly due to two of the site's features (reviewed in [21][22][23][24]).…”

Section: Introductionmentioning

confidence: 99%

Existing drugs as broad-spectrum and potent inhibitors for Zika virus by targeting NS2B-NS3 interaction

et al. 2017

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Recent outbreaks of Zika virus (ZIKV) highlight an urgent need for therapeutics.The protease complex NS2B-NS3 plays essential roles during flaviviral polyprotein processing, and thus represents an attractive drug target. Here, we developed a split luciferase complementation-based high-throughput screening assay to identify orthosteric inhibitors that directly target flavivirus NS2B-NS3 interactions. By screening a total of 2 816 approved and investigational drugs, we identified three potent candidates, temoporfin, niclosamide, and nitazoxanide, as flavivirus NS2B-NS3 interaction inhibitors with nanomolar potencies. Significantly, the most potent compound, temoporfin, not only inhibited ZIKV replication in human placental and neural progenitor cells, but also prevented ZIKV-induced viremia and mortality in mouse models. Structural docking suggests that temoporfin potentially binds NS3 pockets that hold critical NS2B residues, thus inhibiting flaviviral polyprotein processing in a non-competitive manner. As these drugs have already been approved for clinical use in other indications either in the USA or other countries, they represent promising and easily developed therapies for the management of infections by ZIKV and other flaviviruses.

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Structure of the NS2B-NS3 protease from Zika virus after self-cleavage

Cited by 194 publications

References 46 publications

Characterization of the Zika virus two-component NS2B-NS3 protease and structure-assisted identification of allosteric small-molecule antagonists

Characterization of the Zika virus two-component NS2B-NS3 protease and structure-assisted identification of allosteric small-molecule antagonists

Cis autocatalytic cleavage of glycine‐linked Zika virus NS2B‐NS3 protease constructs

Existing drugs as broad-spectrum and potent inhibitors for Zika virus by targeting NS2B-NS3 interaction

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