In vitro synthesis of active recombinant orthoflavivirus nonstructural polyproteins in detergent micelles for biochemical analysis.

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

SUBMITTER: Takahashi H 

PROVIDER: S-EPMC12820207 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature