A sensor for quantification of macromolecular crowding in living cells

Nat Methods. 2015 Mar;12(3):227-9, 1 p following 229. doi: 10.1038/nmeth.3257. Epub 2015 Feb 2.

Abstract

Macromolecular crowding in cells influences processes such as folding, association and diffusion of proteins and polynucleic acids. Direct spatiotemporal readout of crowding would be a powerful approach for unraveling the structure of the cytoplasm and determining the impact of excluded volume on protein function in living cells. Here, we introduce a genetically encodable fluorescence resonance energy transfer (FRET) sensor for quantifying macromolecular crowding and discuss our application of the sensor in bacterial and mammalian cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biosensing Techniques / methods*
  • Calibration
  • Cytoplasm / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fluorescence Resonance Energy Transfer*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Macromolecular Substances / analysis*
  • Macromolecular Substances / metabolism
  • Molecular Imaging / methods*
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Cyan Fluorescent Protein
  • Luminescent Proteins
  • Macromolecular Substances
  • Recombinant Proteins
  • yellow fluorescent protein, Bacteria
  • Green Fluorescent Proteins

Associated data

  • GENBANK/KP399789
  • GENBANK/KP399790