Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein Dynamics in Living Cells01:19

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

Genetic Code Expansion, Enzymatic Modification, and C-Terminal Labeling Enable Facile Production of Highly Modified α-Synuclein.

bioRxiv : the preprint server for biology·2026
Same author

Selective Narrowing of the Bonding Modes of Plasmonic Nanoantennas.

ACS applied materials & interfaces·2026
Same author

Single-molecule fluorescence spectroscopy of fast protein dynamics.

Current opinion in structural biology·2026
Same author

A stochastic mechanism drives fast substrate translocation in the AAA+ machine ClpB.

Nature communications·2026
Same author

Basic Science and Pathogenesis.

Alzheimer's & dementia : the journal of the Alzheimer's Association·2025
Same author

Low-order assemblies drive oncogenic RTK fusion signaling without condensation.

bioRxiv : the preprint server for biology·2025

Related Experiment Video

Updated: Jul 6, 2026

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
11:55

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling

Published on: May 29, 2011

Using fluorescence correlation spectroscopy to study conformational changes in denatured proteins.

Eilon Sherman1, Anna Itkin, Yosef Yehuda Kuttner

  • 1Chemical Physics Department, Weizmann Institute of Science, Rehovot 76100, Israel.

Biophysical Journal
|March 11, 2008
PubMed
Summary
This summary is machine-generated.

Fluorescence correlation spectroscopy accurately measures protein size, even in complex solutions. Studies show denatured proteins expand with increasing denaturant concentration, with variations between protein L and adenylate kinase.

More Related Videos

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
14:04

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Published on: April 25, 2021

Related Experiment Videos

Last Updated: Jul 6, 2026

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
11:55

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling

Published on: May 29, 2011

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
14:04

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Published on: April 25, 2021

Area of Science:

  • Biophysics
  • Protein Chemistry
  • Analytical Chemistry

Background:

  • Fluorescence correlation spectroscopy (FCS) is a sensitive technique for studying biomolecular dynamics and hydrodynamics.
  • Accurate determination of protein size during folding is crucial for understanding protein behavior.
  • Previous studies noted potential artifacts in FCS data analysis affecting size determination.

Purpose of the Study:

  • To validate the accuracy of FCS for measuring diffusion coefficients in solutions with high co-solute concentrations.
  • To investigate the structural changes, specifically expansion, of denatured states of protein L and adenylate kinase (AK) using FCS.
  • To explore potential correlations between protein unfolding and coil-globule transitions in denatured states.

Main Methods:

  • Measurements of diffusion coefficients for small molecules and proteins in solutions with high concentrations of glycerol and guanidinium hydrochloride (GuHCl).
  • Application of FCS to determine the hydrodynamic size of protein L and AK in various denaturant concentrations.
  • Comparison of FCS results with existing data and single-molecule fluorescence histograms.

Main Results:

  • FCS accurately determines diffusion coefficients, validating the Stokes-Einstein relation even in highly concentrated glycerol and GuHCl solutions.
  • The denatured state of protein L shows significant expansion with increasing GuHCl concentration, resembling a polymer's coil-globule transition.
  • The denatured state of AK exhibits limited expansion beyond 2 M GuHCl, aligning with other experimental observations.

Conclusions:

  • FCS is a reliable method for probing biomolecular size and dynamics, even in challenging solution conditions.
  • Denatured protein structures exhibit varying degrees of expansion, suggesting distinct coil-globule transition behaviors.
  • A potential correlation between unfolding transitions and coil-globule transitions is suggested for AK due to similar denaturant concentration dependencies.