Phanta: a non-fluorescent photochromic acceptor for pcFRET

PLoS One. 2013 Sep 30;8(9):e75835. doi: 10.1371/journal.pone.0075835. eCollection 2013.

Abstract

We have developed an orange non-fluorescent photochromic protein (quantum yield, 0.003) we call Phanta that is useful as an acceptor in pcFRET applications. Phanta can be repeatedly inter-converted between the two absorbing states by alternate exposure to cyan and violet light. The absorption spectra of Phanta in one absorbing state shows excellent overlap with the emission spectra of a number of donor green fluorescent proteins including the commonly used EGFP. We show that the Phanta-EGFP FRET pair is suitable for monitoring the activation of caspase 3 in live cells using readily available instrumentation and a simple protocol that requires the acquisition of two donor emission images corresponding to Phanta in each of its photoswitched states. This the first report of a genetically encoded non-fluorescent acceptor for pcFRET.

MeSH terms

  • Amino Acid Sequence
  • Biosensing Techniques / methods*
  • Fluorescence Resonance Energy Transfer / methods*
  • Green Fluorescent Proteins / genetics*
  • Green Fluorescent Proteins / metabolism
  • Molecular Sequence Data
  • Photochemistry / methods*
  • Protein Engineering
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Recombinant Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins

Grants and funding

The authors have no support or funding to report.