Quantitative tracking of protein trafficking to the nucleus using cytosolic protein delivery by nanoparticle-stabilized nanocapsules

Bioconjug Chem. 2015 Jun 17;26(6):1004-7. doi: 10.1021/acs.bioconjchem.5b00141. Epub 2015 Jun 2.

Abstract

We describe a method for quantitative monitoring of subcellular protein trafficking using nanoparticle-stabilized nanocapsules for protein delivery. This method provides rapid delivery of the protein into the cytosol, eliminating complications from protein homeostasis processes found with cellularly expressed proteins. After delivery, nuclear protein trafficking was followed by real time microscopic imaging. Quantitative analyses of the accumulation percentage and the import dynamics of the nuclear protein trafficking, demonstrate the utility of this method for studying intracellular trafficking systems.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Nucleus / metabolism*
  • Cytosol / metabolism*
  • Fluorescent Dyes / administration & dosage
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / pharmacokinetics
  • Green Fluorescent Proteins / administration & dosage*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / pharmacokinetics
  • HeLa Cells
  • Humans
  • Models, Molecular
  • Nanocapsules / chemistry*
  • Nanoparticles / chemistry*
  • Nuclear Localization Signals
  • Optical Imaging
  • Protein Transport
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / pharmacokinetics

Substances

  • Fluorescent Dyes
  • Nanocapsules
  • Nuclear Localization Signals
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins