Jove
Visualize
Contact Us

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

Single-molecule dissection of CFTR folding defects and pharmacological rescue.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Snapshots of the dynamic basis of NTSR1 G protein subtype promiscuity.

Nature·2026
Same author

Non-equilibrium snapshots of ligand efficacy at the μ-opioid receptor.

Nature·2025
Same author

Parallel stopped-flow interrogation of diverse biological systems at the single-molecule scale.

Nature methods·2025
Same author

Metabolic Flexibility of Microglia: Energy Substrate Utilization and Impact on Neuronal Metabolism.

Journal of neurochemistry·2025
Same author

Structurally diverse viral inhibitors converge on a shared mechanism to stall the antigen transporter TAP.

Proceedings of the National Academy of Sciences of the United States of America·2025
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 Experiment Video

Updated: Jun 10, 2026

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae
15:12

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae

Published on: May 10, 2014

14.6K

CFTR function, pathology and pharmacology at single-molecule resolution.

Jesper Levring1, Daniel S Terry2, Zeliha Kilic2

  • 1Laboratory of Membrane Biology and Biophysics, The Rockefeller University, New York, NY, USA.

Nature
|March 23, 2023
PubMed
Summary
This summary is machine-generated.

Cystic fibrosis transmembrane conductance regulator (CFTR) NBDs dimerize before opening, controlling chloride flow. This mechanism explains how CFTR potentiators work and informs new cystic fibrosis therapies.

More Related Videos

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
09:59

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Published on: March 9, 2015

12.3K
Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
09:59

Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking

Published on: March 16, 2017

8.9K

Related Experiment Videos

Last Updated: Jun 10, 2026

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae
15:12

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae

Published on: May 10, 2014

14.6K
Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
09:59

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Published on: March 9, 2015

12.3K
Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
09:59

Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking

Published on: March 16, 2017

8.9K

Area of Science:

  • Biophysics
  • Molecular Biology
  • Ion Channel Physiology

Background:

  • The cystic fibrosis transmembrane conductance regulator (CFTR) is a crucial anion channel regulating epithelial salt and fluid balance.
  • Dysfunction of CFTR leads to cystic fibrosis, a severe, incurable genetic disorder.
  • Previous studies analyzed CFTR's electrophysiology and determined its structure in distinct conformations, but lacked direct functional correlations.

Purpose of the Study:

  • To elucidate the gating mechanism of the human CFTR channel.
  • To correlate CFTR's structure with its function in regulating ion transport.
  • To understand how disease-causing mutations and potentiator drugs affect CFTR function.

Main Methods:

  • Ensemble functional measurements
  • Single-molecule fluorescence resonance energy transfer (smFRET)
  • Electrophysiology
  • Kinetic simulations

Main Results:

  • The two nucleotide-binding domains (NBDs) of CFTR dimerize prior to channel opening.
  • CFTR operates via an allosteric gating mechanism involving NBD dimerization and ATP hydrolysis.
  • Potentiators like ivacaftor increase channel activity by enhancing pore opening during NBD dimerization.
  • Disease-associated mutations (G551D, L927P) impair NBD dimerization efficiency.

Conclusions:

  • A detailed gating mechanism for CFTR has been proposed, involving NBD dimerization as a key step.
  • Understanding this mechanism provides insights into the action of potentiator drugs.
  • These findings pave the way for developing more effective therapeutic strategies for cystic fibrosis.