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Related Experiment Video

Updated: Feb 24, 2026

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
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mSumireF a Monomeric Violet Fluorescent Protein.

Jacob Kress1, Sookyeong Kim1, Caitlynn Bryant1

  • 1Department of Chemistry & Biochemistry Program, Wellesley College, 106 Central Street, Wellesley, Massachusetts 02481, United States.

ACS Bio & Med Chem Au
|February 23, 2026
PubMed
Summary

Researchers developed mSumireF, a brighter, less-aggregating violet fluorescent protein. This improved protein is ideal for multicolor biological imaging and FRET biosensors, expanding options for researchers.

Keywords:
Sumirehetero-FRET donorhomo-FRETmonomeric violet fluorescent proteinoligomerization

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biophysics

Background:

  • Genetically encoded fluorescent proteins are essential tools for multicolor biological imaging.
  • Developing well-behaved fluorescent proteins with distinct spectral properties is crucial for advancing multicolor experiments.
  • Existing violet fluorescent proteins can suffer from dimerization and reduced brightness, limiting their utility.

Purpose of the Study:

  • To engineer a monomeric variant of the violet fluorescent protein Sumire with improved brightness and reduced oligomerization.
  • To characterize the biophysical properties of the engineered mSumireF variant.
  • To evaluate the utility of mSumireF in FRET-based biosensor applications.

Main Methods:

  • Site-directed mutagenesis was used to engineer the mSumireF variant.
  • Oligomerization was assessed using the organized smooth endoplasmic reticulum (OSER) assay in mammalian cells and solution-based assays.
  • Fluorescence properties including brightness and spectral characteristics were measured.
  • Förster Resonance Energy Transfer (FRET) efficiency was evaluated using mSumireF as a donor with various acceptor fluorescent proteins.

Main Results:

  • The mSumireF variant, engineered with a specific amino acid reversion, restored brightness lost in a previous monomeric variant.
  • mSumireF demonstrated significantly reduced oligomerization tendency compared to the original Sumire and other monomeric variants.
  • mSumireF effectively functioned as a FRET donor when paired with mCerulean3, mTurquoise2, and LSSmScarlet.
  • The engineered protein maintained desirable spectral properties for multicolor applications.

Conclusions:

  • mSumireF is a bright, monomeric violet fluorescent protein that overcomes limitations of previous variants.
  • This improved fluorescent protein expands the available palette for multicolor imaging and reduces concerns about protein aggregation.
  • mSumireF is suitable for constructing advanced FRET-based biosensors, including those for ATP detection.