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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Protein Diffusion in the Membrane01:24

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Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...
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Updated: Sep 15, 2025

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT
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Surface-Tethering Enhances Precision in Measuring Diffusion Within 3D Protein Condensates.

Emily R Sumrall1,2, Guoming Gao1,2,3, Shelby Stakenas4

  • 1Biophysics Graduate Program, University of Michigan, Ann Arbor, MI 48109, USA.

Biorxiv : the Preprint Server for Biology
|July 16, 2025
PubMed
Summary
This summary is machine-generated.

Surface tethering immobilizes biomolecular condensates, preventing artifacts from their natural motion. This method ensures accurate measurements of molecular diffusion within these essential cellular structures, crucial for understanding their function.

Keywords:
FUSRNAbiomolecular condensatesdiffusionmembraneless organellessingle-molecule trackingsurface passivation

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Area of Science:

  • Cell biology
  • Biophysics
  • Molecular dynamics

Background:

  • Biomolecular condensates (membraneless organelles) are vital for cellular organization and function.
  • Studying their internal dynamics requires advanced imaging techniques, but condensate motion can cause measurement errors.

Purpose of the Study:

  • To investigate and mitigate artifacts caused by condensate Brownian motion in diffusion measurements.
  • To establish a robust method for accurate quantification of molecular dynamics within condensates.

Main Methods:

  • Utilized three surface-tethering strategies (DNA, protein, antibody) to immobilize FUS protein condensates.
  • Employed super-resolved imaging and single-molecule tracking to analyze molecular mobility.
  • Conducted simulations to model condensate behavior and diffusion across physiological parameters.

Main Results:

  • Surface tethering effectively suppressed condensate Brownian motion without altering their liquid-like properties.
  • Untethered condensates led to inaccurate diffusion measurements, especially for structured RNAs.
  • Tethering strategies offered tunable control over condensate stability and dynamics.

Conclusions:

  • Surface tethering is essential for accurate quantification of intra-condensate molecular dynamics.
  • This approach provides a reliable methodological framework for studying membraneless organelles.
  • Guidelines were developed to help researchers determine the necessity and extent of tethering.