Modification of CCMV nanocages for enzyme encapsulation

Lise Schoonen, Jan C.M. van Hest

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

1 Citation (Scopus)

Abstract

In cellular systems, compartmentalization plays an important role in the protection and regulation of enzymes. Controlled encapsulation of enzymes in nanocompartments is crucial in understanding biocatalytic processes in the cellular environment. We have recently described an enzymatic method to covalently attach enzymes, equipped with a small recognition peptide, to the interior of viral capsids. Viral capsids are especially interesting in this respect, as they form very well-defined nanoparticles with a uniform size and shape. Here, we describe the relevant experimental procedures to encapsulate a model enzyme into the interior of a viral capsid, purify the resulting viral capsids, and measure the catalytic activity of the encapsulated enzymes.

Original languageEnglish
Title of host publicationProtein Scaffolds : Design, Synthesis, and Applications
EditorsA.K. Udit
Place of PublicationDordrecht
PublisherSpringer
Pages69-83
Number of pages15
ISBN (Electronic)978-1-4939-7893-9
ISBN (Print)978-1-4939-7892-2
DOIs
Publication statusPublished - 5 Jun 2018

Publication series

NameMethods in Molecular Biology
Volume1798
ISSN (Print)1064-3745

Keywords

  • Candida antarctica lipase B
  • Cowpea chlorotic mottle virus
  • Esterase activity
  • Self-assembly
  • Sortagging
  • Sortase A
  • Viral capsid
  • Gene Expression
  • Models, Molecular
  • Aminoacyltransferases/chemistry
  • Virus Assembly
  • Bromovirus
  • Capsid Proteins/chemistry
  • Nanocapsules/chemistry
  • Chromatography, Affinity
  • Enzymes, Immobilized/chemistry
  • Capsid/chemistry
  • Mass Spectrometry
  • Protein Conformation
  • Bacterial Proteins/chemistry
  • Cysteine Endopeptidases/chemistry
  • Recombinant Fusion Proteins/chemistry
  • Hydrogen-Ion Concentration

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