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Researchers studied the evolutionary history of fluorescent proteins to understand red emission. They found that the mPlum protein

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Area of Science:

  • Biophysics
  • Photochemistry
  • Protein Engineering

Background:

  • Solvent reorganization around excited chromophores causes emission shifts.
  • Wild-type green fluorescent protein lacks this solvation response due to environmental rigidity.
  • This rigidity prevents nonradiative decay, maintaining fluorescence efficiency.

Purpose of the Study:

  • Investigate the dynamic Stokes shift in fluorescent proteins through their evolutionary history.
  • Understand the structural and environmental factors enabling solvation response in red-emitting variants like mPlum.
  • Identify the molecular determinants responsible for the picosecond solvation response in mPlum.

Main Methods:

  • Time-resolved fluorescence spectroscopy was employed.
  • Studied the evolutionary trajectory of fluorescent proteins, including mPlum.
  • Analyzed emission shifts across different temperatures and evolutionary stages.

Main Results:

  • The red-emitting fluorescent protein mPlum exhibits a picosecond solvation response above its glass transition temperature.
  • This dynamic Stokes shift is absent in evolutionary ancestors of mPlum.
  • A single amino acid residue near the chromophore is implicated as the cause of the solvation response.

Conclusions:

  • Evolutionary selection for red emission in mPlum has resulted in a chromophore environment that permits solvent reorganization.
  • This reorganization is responsible for the observed picosecond solvation response and far-red emission.
  • Structural modifications, particularly a single residue, are key to enabling this photophysical behavior.