Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Studying the Cytoskeleton

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Spatiotemporal fluctuations in fluorescence intensity of rhodamine phalloidin-labeled actin filaments.

The Journal of biological chemistry·2025

Same author

Multiple Mechanisms to Regulate Actin Functions: "Fundamental" Versus Lineage-Specific Mechanisms and Hierarchical Relationships.

Biomolecules·2025

Same author

Ras activation by hydrostatic pressure involves GDP release and is enhanced by GAP and GEF in vitro.

Archives of biochemistry and biophysics·2025

Same author

Nestin Forms a Flexible Cytoskeleton by Means of a Huge Tail Domain That Is Reversibly Stretched and Contracted by Weak Forces.

Cells·2025

Same author

A simple model and rules for the evolution of microbial mutualistic symbiosis with positive fitness feedbacks.

Theoretical population biology·2024

Same author

CHLOROPLAST UNUSUAL POSITIONING 1 is a plant-specific actin polymerization factor regulating chloroplast movement.

The Plant cell·2023

Search research articles

Related Experiment Video

Updated: Jun 28, 2026

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Published on: February 4, 2021

Visualizing myosin-actin interaction with a genetically-encoded fluorescent strain sensor.

Sosuke Iwai¹, Taro Q P Uyeda

¹Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan. sosuke@aist.go.jp

Proceedings of the National Academy of Sciences of the United States of America

|October 31, 2008

Summary

Researchers developed a novel fluorescent strain sensor to visualize force-induced protein conformational changes within living cells. This sensor, based on proximity imaging, successfully detected myosin-actin interactions and conformational shifts in response to cellular stress.

More Related Videos

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers

Published on: May 4, 2022

Tuning the Contractility and Deformation Modes of Active Actin-Based Assemblies In Vitro: From Two-Dimensional Active Networks to Liquid Crystal Drops

Tuning the Contractility and Deformation Modes of Active Actin-Based Assemblies In Vitro: From Two-Dimensional Active Networks to Liquid Crystal Drops

Published on: July 11, 2025

Related Experiment Videos

Last Updated: Jun 28, 2026

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Published on: February 4, 2021

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers

Published on: May 4, 2022

Tuning the Contractility and Deformation Modes of Active Actin-Based Assemblies In Vitro: From Two-Dimensional Active Networks to Liquid Crystal Drops

Tuning the Contractility and Deformation Modes of Active Actin-Based Assemblies In Vitro: From Two-Dimensional Active Networks to Liquid Crystal Drops

Published on: July 11, 2025

Area of Science:

Biophysics
Molecular Cell Biology
Biochemistry

Background:

Force-induced protein conformational changes are well-documented in vitro, but their occurrence in vivo remains largely uncharacterized.
Understanding these in vivo changes is crucial for comprehending cellular mechanics and protein function under physiological conditions.

Purpose of the Study:

To develop and validate a genetically encoded fluorescent sensor for detecting force-induced protein conformational changes (strains) within living cells.
To investigate the conformational states of myosin-actin interactions in Dictyostelium cells under varying conditions.

Main Methods:

Development of a PRIM-based strain sensor module (PriSSM) using tandem fusion of normal and circularly permuted GFP.
Insertion of PriSSM into Dictyostelium myosin II motor domains to detect conformational changes upon F-actin binding.
Utilized spectral changes in GFP moieties to report on the proximity and strain status of PriSSM.

Main Results:

The PriSSM sensor successfully detected conformational changes in myosin II motor domains upon binding to F-actin.
Mutations within the myosin motor domain were found to influence the lever arm position in the rigor state.
Visualized myosin II-F-actin interactions in Dictyostelium cells, revealing increased binding under ATP depletion or hyperosmotic stress.

Conclusions:

The PRIM-based strain sensor (PriSSM) provides a powerful tool for real-time visualization of force-induced protein conformational changes in vivo.
This technology offers a generalizable approach for studying mechanobiology and protein dynamics within living cells.