ArticleNpj viruses2026
In vitro synthesis of active recombinant orthoflavivirus nonstructural polyproteins in detergent micelles for biochemical analysis.
Article in Npj viruses, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The nonstructural (NS) proteins of orthoflaviviruses are cleaved from a precursor polyprotein by both host and viral proteases, subsequently forming a replication complex. However, the biochemical properties underlying polyprotein cleavage and individual NS proteins within the complex remain largely unexplored. Here, we synthesized recombinant dengue virus (DENV-2) polyproteins containing all NS proteins for the biochemical evaluation of the synthesis and cleavage of the polyproteins. The results showed successful synthesis of the full-length polyprotein, which efficiently underwent self-cleavage when the detergent n-dodecyl-β-maltoside (DDM) was added. In addition, the cleaved NS proteins formed a complex in the DDM micelle and possessed the enzymatic activities of NS3 and NS5. Leveraging this in vitro system, we found that polyprotein was cleaved predominantly via cis-intramolecular activity, immediately following polyprotein synthesis. Therefore, the orthoflavivirus polyproteins synthesized here is potentially a valuable tool for the biochemical analysis of orthoflavivirus polyproteins.
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