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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

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Modulation of Fluorescent Protein Chromophores To Detect Protein Aggregation with Turn-On Fluorescence

Yu Liu, Charles H Wolstenholme, Gregory C Carter

  • 1Department of Chemistry , University of Washington , Seattle , Washington 98105 , United States.

Journal of the American Chemical Society
|June 9, 2018
PubMed

Insights

We developed a new fluorescent method to visualize misfolded proteins in live cells. This technique detects previously invisible misfolded soluble proteins, offering new insights into protein aggregation and cellular health.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Biophysics

Background:

  • Protein misfolding and aggregation are implicated in various diseases.
  • Current methods struggle to detect soluble misfolded proteins in live cells.

Purpose of the Study:

  • To develop a novel fluorogenic method for visualizing specific protein misfolding and aggregation in live cells.
  • To enable the detection of previously undetectable misfolded soluble proteins.

Main Methods:

  • Utilized structurally modulated fluorescent protein chromophores.
  • Combined photophysical analysis, X-ray crystallography, and theoretical calculations.
  • Employed bioorthogonal conjugation of fluorophores to Halo-tag fused proteins.

Main Results:

  • Demonstrated that fluorescence is triggered by the inhibition of twisted-intramolecular charge transfer in rigid environments.
  • Successfully visualized both misfolded and aggregated protein species in live cells.
  • Showcased the ability to detect previously invisible misfolded soluble proteins.

Conclusions:

  • The developed fluorogenic method provides a powerful tool for studying protein conformational changes in real-time.
  • This approach offers unprecedented sensitivity for detecting misfolded proteins, advancing our understanding of proteinopathies.
  • Represents the first application of fluorescent protein chromophores for detecting protein conformational collapse in live cellular systems.

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