Single-Molecule Specific Mislocalization of Red Fluorescent Proteins in Live Escherichia coli

Biophys J. 2016 Jul 12;111(1):25-7. doi: 10.1016/j.bpj.2016.05.047. Epub 2016 Jun 20.

Abstract

Tagging of individual proteins with genetically encoded fluorescent proteins (FPs) has been used extensively to study localization and interactions in live cells. Recent developments in single-molecule localization microscopy have enabled the dynamic visualization of individual tagged proteins inside living cells. However, tagging proteins with FPs is not without problems: formation of insoluble aggregates and inhibition of native functions of the protein are well-known issues. Previously reported artifacts manifest themselves at all expression levels of the FP-tagged proteins, making the design of control experiments relatively straightforward. Here, we describe a previously uncharacterized mislocalization artifact of Entacmaea quadricolor red fluorescent protein variants that is detectable at the single-molecule level in live Escherichia coli cells.

MeSH terms

  • Escherichia coli / metabolism*
  • Escherichia coli / physiology
  • Luminescent Proteins / metabolism*
  • Microbial Viability*
  • Protein Transport
  • Red Fluorescent Protein
  • Single Molecule Imaging*

Substances

  • Luminescent Proteins