Expression and Purification of Vaccinia Virus DNA Topoisomerase IB Produced in the Silkworm–Baculovirus Expression System

Xu, Jian; Lee, Jae Man; Tatsuke, Tsuneyuki; Ebihara, Takeru; Masuda, Akihiro; Hino, Masato; Morokuma, Daisuke; Fujita, Ryosuke; Mon, Hiroaki; Kusakabe, Takahiro; Takahashi, Masateru

doi:10.1007/s12033-019-00184-4

Cited by 6 publications

(4 citation statements)

References 41 publications

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“…Nitroxoline exerts its antibacterial effects by chelating metal ions including Fe2 + , Mn2 + , and Mg2 + . 53 Although poxviruses depend on the availability of bivalent cations for effective replication, 54,55 the antiviral activity of nitroxoline was not affected by the addition of metal ions. This shows that nitroxoline's antiviral and antibacterial mechanisms of action differ substantially.…”

Section: Discussionmentioning

confidence: 99%

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Bojkova

Zöller

Tietgen

et al. 2023

Journal of Medical Virology

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The antiviral drugs tecovirimat, brincidofovir, and cidofovir are considered for mpox (monkeypox) treatment despite a lack of clinical evidence. Moreover, their use is affected by toxic side‐effects (brincidofovir, cidofovir), limited availability (tecovirimat), and potentially by resistance formation. Hence, additional, readily available drugs are needed. Here, therapeutic concentrations of nitroxoline, a hydroxyquinoline antibiotic with a favourable safety profile in humans, inhibited the replication of 12 mpox virus isolates from the current outbreak in primary cultures of human keratinocytes and fibroblasts and a skin explant model by interference with host cell signalling. Tecovirimat, but not nitroxoline, treatment resulted in rapid resistance development. Nitroxoline remained effective against the tecovirimat‐resistant strain and increased the anti‐mpox virus activity of tecovirimat and brincidofovir. Moreover, nitroxoline inhibited bacterial and viral pathogens that are often co‐transmitted with mpox. In conclusion, nitroxoline is a repurposing candidate for the treatment of mpox due to both antiviral and antimicrobial activity.

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Section: Discussionmentioning

confidence: 99%

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Bojkova

Zöller

Tietgen

et al. 2023

Journal of Medical Virology

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show abstract

“…Nitroxoline exerts its antibacterial effects by chelating metal ions including Fe2 + , Mn2 + , and Mg2 + [Pelletier et al, 1995]. Although poxviruses depend on the availability of bivalent cations for effective replication [Li et al, 2016; Xu J et al, 2019], the antiviral activity of nitroxoline was not affected by the addition of metal ions. This shows that nitroxoline’s antiviral and antibacterial mechanisms of action differ substantially.…”

Section: Discussionmentioning

confidence: 99%

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Bojkova

Zöller

Tietgen

et al. 2022

Preprint

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The antiviral drugs tecovirimat, brincidofovir, and cidofovir are considered for mpox (monkeypox) treatment despite a lack of clinical evidence. Moreover, their use is affected by toxic side-effects (brincidofovir, cidofovir), limited availability (tecovirimat), and potentially by resistance formation. Hence, additional, readily available drugs are needed. Here, therapeutic concentrations of nitroxoline, a hydroxyquinoline antibiotic with a favourable safety profile in humans, inhibited the replication of 12 mpox virus isolates from the current outbreak in primary cultures of human keratinocytes and fibroblasts and a skin explant model by interference with host cell signalling. Tecovirimat, but not nitroxoline, treatment resulted in rapid resistance development. Nitroxoline remained effective against the tecovirimat-resistant strain and increased the anti-mpox virus activity of tecovirimat and brincidofovir. Moreover, nitroxoline inhibited bacterial and viral pathogens that are often co-transmitted with mpox. In conclusion, nitroxoline is a repurposing candidate for the treatment of mpox due to both antiviral and antimicrobial activity.

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“…It has been demonstrated that the ST/SC-based protein cyclization (SpyRing) of certain POIs (case-by-case), such as beta-lactamase and dihydrofolate reductase, confers higher thermal stability and activity [15,35,36]. Since there has been already some cases that recombinant proteins from silkworm-BEVS hold better stability and activity, further incorporation of SpyRing system might have a synergistic effect [37,38]. We are now challenging the silkworm-SpyRing-BEVS using some POIs in our laboratory.…”

Section: Discussionmentioning

confidence: 99%

Development of SpyTag/SpyCatcher-Bacmid Expression Vector System (SpyBEVS) for Protein Bioconjugations Inside of Silkworms

Kato

Park

2019

IJMS

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Protein conjugations at post-translational levels are known to be essential to protein stability and function. Recently, it has been proven that the split protein CnaB2 (SpyTag/SpyCatcher, ST/SC) from Streptococcus pyogenes can induce covalent conjugation rapidly and efficiently under various conditions. The protein of interest fused with the split protein SC/ST could be assembled spontaneously. In light of this finding, we introduced the ST/SC protein coupling concept into the silkworm-bacmid protein expression system (SpyBEVS). As a proof of concept, we first examined and confirmed that a competent ligation occurred between ST/SC-fused protein partners in vitro in cultured silkworm cells and in vivo in silkworm larvae by co-infection of several recombinant baculoviruses. The protein conjugation could be also achieved sufficiently by a simple one-step mixture of purified ST/SC-tagged peptide-protein pairs in vitro. Given the flexibility and robustness of silkworm-BEVS, our results on SpyBEVS show an alternative method for enabling the production of protein decorations in vitro and inside of silkworms.

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Expression and Purification of Vaccinia Virus DNA Topoisomerase IB Produced in the Silkworm–Baculovirus Expression System

Cited by 6 publications

References 41 publications

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Repurposing of the antibiotic nitroxoline for the treatment of mpox

Development of SpyTag/SpyCatcher-Bacmid Expression Vector System (SpyBEVS) for Protein Bioconjugations Inside of Silkworms

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