Colorful virus-like particles: fluorescent protein packaging by the Qβ capsid

Biomacromolecules. 2011 Nov 14;12(11):3977-81. doi: 10.1021/bm200983k. Epub 2011 Oct 13.

Abstract

Qβ virus-like particles encapsulating multiple copies of fluorescent proteins were generated in high yields using a modular system enhanced by specific engineered RNA--protein interactions. The resulting particles were structurally indistinguishable from recombinant Qβ alone. The encapsidated proteins were nearly identical in photochemical properties to monomeric analogues, were more stable toward thermal degradation, and were protected from proteolytic cleavage. Residues on the outer capsid surface were chemically derivatized by acylation and azide--alkyne cycloaddition without affecting the fluorescence properties of the packaged proteins. A high-affinity carbohydrate-based ligand of the CD22 receptor was thereby attached, and specific cell labeling by the particles was successfully detected by flow cytometry and confocal laser microscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allolevivirus*
  • Animals
  • Aptamers, Nucleotide / metabolism
  • CHO Cells
  • Capsid / metabolism*
  • Capsid Proteins / metabolism*
  • Cloning, Molecular
  • Cricetinae
  • Green Fluorescent Proteins / metabolism*
  • Humans
  • Luminescent Proteins / metabolism*
  • Protein Stability
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism*
  • Sialic Acid Binding Ig-like Lectin 2 / metabolism
  • Virus Attachment

Substances

  • Aptamers, Nucleotide
  • CD22 protein, human
  • Capsid Proteins
  • Cyan Fluorescent Protein
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Sialic Acid Binding Ig-like Lectin 2
  • blue fluorescent protein, Aequorea victoria
  • Green Fluorescent Proteins