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Related Concept Videos

Actin and Myosin in Muscle Contraction01:16

Actin and Myosin in Muscle Contraction

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Actin and myosin are contractile proteins that form the sarcomere found in skeletal muscle tissues for regulating muscle contraction. Actin, a globular contractile protein, interacts with myosin for muscle contraction. The skeletal tissue appears striped or striated under a microscope due to the repeated arrangement of contractile proteins actin and myosin along the length of myofibrils. Dark A bands and light I bands repeat along myofibrils, and the alignment of myofibrils in the cell causes...
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The Role of Actin and Myosin in Non-muscle Cells01:10

The Role of Actin and Myosin in Non-muscle Cells

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Actin and myosin or actomyosin filaments also play a significant role in cells other than those involved in muscle contraction (which occurs within the sarcomere of muscle cells). The mechanism of non-muscle cell contractile bundles was first observed in Dictyostelium and Acanthamoeba. In non-muscle cells, two bundles are commonly found: stress fibers and actomyosin adherence belts. These contractile bundles are smaller and less organized than the ones found in muscle cells. They  are held...
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Actin Treadmilling01:18

Actin Treadmilling

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Actin filaments undergo polymerization and depolymerization from either end. The polymerization and depolymerization rates depend on the cytosolic concentration of free G-actins. The polymerization rate is generally higher at the plus or barbed end, while the depolymerization rate is higher at the minus or pointed end. At a steady state, critical concentration describes the concentration of free G-actin monomers at which the polymerization rate at the plus end is equal to that of the...
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Overview of Myosin Structure and Function01:15

Overview of Myosin Structure and Function

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Myosins are a family of molecular motor proteins, first identified in the skeletal muscles, where they are responsible for muscle contraction. Along with their role in muscle contraction, these proteins also play a role in the intracellular transport of molecules and vesicles. There are twenty-four classes of myosins based on their domain sequence and organization. Of the twenty-four, six classes (Myosin I, Myosin II, Myosin V, Myosin VI, Myosin VII, and Myosin X)  have been well...
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Introduction to Actin01:26

Introduction to Actin

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Actin is a highly conserved cytoskeletal protein found abundantly in eukaryotic cells. It constitutes 10% weight of the total cellular protein in muscle cells, while in non-muscle cells, it is lower and makes up around 1–5 percent of the total cell protein. Actin found in the unicellular amoebae and complex multicellular animals is around 80% similar, demonstrating their conservation over a billion years of evolution.  Actin coding genes are conserved within species and across...
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Actin Polymerization01:42

Actin Polymerization

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Actin polymerization occurs through the head-to-tail association of binding sites on monomeric actin or G-actin to form filamentous or F-actin. The polymerization can be divided into three phases ̶  nucleation, elongation, and steady-state phase.
The nucleation phase involves forming a stable nucleus consisting of three actin monomers to form a new actin filament. Actin-binding proteins such as formins and Arp2/3 complex help filament growth post-nucleation. The Formins form straight...
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Related Experiment Video

Updated: Feb 9, 2026

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
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Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin

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High-throughput screen, using time-resolved FRET, yields actin-binding compounds that modulate actin-myosin structure

Piyali Guhathakurta1, Ewa Prochniewicz1, Benjamin D Grant2

  • 1Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.

The Journal of Biological Chemistry
|June 6, 2018
PubMed
Summary

Researchers developed a new time-resolved Förster Resonance Energy Transfer (TR-FRET) assay to find drugs that modify actin-myosin interactions, crucial for muscle function and movement.

Keywords:
actinfluorescence resonance energy transfer (FRET)high-throughput screening (HTS)myosinpeptide

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Last Updated: Feb 9, 2026

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Measuring Protein Binding to F-actin by Co-sedimentation
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Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers
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Area of Science:

  • Biochemistry and Molecular Biology
  • Cellular Mechanics
  • Drug Discovery

Background:

  • Actin-myosin interactions are fundamental for cellular force generation and movement.
  • Disruptions in actin-myosin function due to genetic or post-translational changes lead to severe muscle disorders.
  • Targeting the actin-myosin interface offers a therapeutic strategy for muscle diseases.

Purpose of the Study:

  • To develop and validate a novel time-resolved Förster Resonance Energy Transfer (TR-FRET) biosensor for detecting modulators of actin-myosin interactions.
  • To identify small molecules that bind to the actin-myosin interface and alter the structural dynamics of the complex.
  • To establish a high-throughput screening method for discovering allosteric modulators of actomyosin function.

Main Methods:

  • Utilized a TR-FRET assay by labeling actin at Cys-374 with a fluorescent donor and a peptide mimicking myosin's essential light chain N-terminus with an acceptor.
  • Validated the biosensor by confirming overlapping binding sites of the peptide and myosin on actin and observing decreased FRET upon myosin binding.
  • Performed a high-throughput screen of 727 small molecules using fluorescence lifetime measurements.

Main Results:

  • Identified 10 compounds that significantly affected FRET in the micromolar range.
  • Observed that most identified compounds modulated actin-activated myosin ATPase activity.
  • Detected alterations in actin's microsecond dynamics using transient phosphorescence anisotropy.

Conclusions:

  • The developed actin-ANT TR-FRET assay is effective for discovering pharmacologically active compounds impacting actin dynamics and actomyosin function.
  • This assay demonstrates the feasibility of identifying allosteric modulators of the actin-myosin interaction.
  • The findings pave the way for developing novel therapies for muscle disorders by targeting actomyosin regulation.