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TitleAn engineered closed-shell, two-component, 480-subunit nucleocapsid.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 123, Issue 22, Page e2530090123, Year 2026
Publish dateJun 2, 2026
AuthorsMikail D Levasseur / Naohiro Terasaka / Angela Steinauer / Stephan Tetter / Sara Pfister / Beat H Meier / Donald Hilvert /
PubMed AbstractSelf-assembling protein cages are valuable nanoscale containers for biotechnology and medical applications. Two-component systems are especially attractive due to their potential for functional ...Self-assembling protein cages are valuable nanoscale containers for biotechnology and medical applications. Two-component systems are especially attractive due to their potential for functional complexity. In this study, we demonstrate that the subunits of the 240-subunit nucleocapsid NC-4, which was previously evolved to package and protect its encoding mRNA, can be split into two fragments without disrupting cage assembly or structure, generating a two-component, 480-subunit capsid. This modification introduces additional termini on the cage's exterior surface, creating opportunities for functionalization. We exploited these new sites by genetically appending peptide and protein tags to the exterior surface of split NC-4 (spNC-4), enabling site-specific glycosylation via posttranslational modification and cell-specific delivery by targeted antibody recruitment. Our findings broaden the utility of the NC-4 nucleocapsid. By extension, splitting related protein compartments that bind diverse cargoes could offer a robust platform for biotechnological applications requiring simultaneous encapsulation and customizable surface modification.
External linksProc Natl Acad Sci U S A / PubMed:42201976 / PubMed Central
MethodsEM (single particle)
Resolution3.1 Å
Structure data

56772

28ro

EMDB-56772, PDB-28ro:
split variant of Aquifex aeolicus lumazine synthase-derived nucleocapsid variant NC-4
Method: EM (single particle) / Resolution: 3.1 Å

Source
  • Escherichia virus lambda
  • aquifex aeolicus (bacteria)
KeywordsVIRUS LIKE PARTICLE / capsid / design / virus mimic

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