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

Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

8.8K
Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
8.8K

You might also read

Related Articles

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

Sort by
Same author

Genome evolution in plant pathogenic bacteria.

Genome biology and evolution·2026
Same author

Ion Channel Dysfunction and Therapeutic Targeting in Salivary Gland Disorders.

Oral diseases·2026
Same author

Advancing the microalgal blue bioeconomy through technological and analytical integration.

Preparative biochemistry & biotechnology·2026
Same author

Nitrogen-Doped Carbon Quantum Dots for FRET-Assisted and Smartphone-Enabled Fluorescent Detection of Melatonin in Biological Samples.

Journal of fluorescence·2026
Same author

Correction: Barriers and Facilitators in the Implementation of the Systematic Medical Appraisal, Referral, and Treatment (SMART) Mental Health Digital Intervention in Rural India: Mixed Methods Process Evaluation Study.

JMIR mental health·2026
Same author

Need for standardization in granulocyte transfusion therapy: Ethical, operational and dose considerations.

Transfusion clinique et biologique : journal de la Societe francaise de transfusion sanguine·2026

Related Experiment Video

Updated: Sep 27, 2025

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
09:32

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development

Published on: June 15, 2017

8.8K

Optogenetic Control of PIP2 Interactions Shaping ENaC Activity.

Tarek Mohamed Abd El-Aziz1,2, Amanpreet Kaur3, Mark S Shapiro1

  • 1Department of Cellular and Integrative Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78228, USA.

International Journal of Molecular Sciences
|April 12, 2022
PubMed
Summary

Phosphatidylinositol 4,5-bisphosphate (PIP2) binding sites on epithelial sodium channel (ENaC) subunits are crucial for channel activity. Mutations in these sites abolish ENaC

Keywords:
CRY2CoroNa GreenENaCPIP2optogeneticphosphoinositidessodium channel

More Related Videos

Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation
08:00

Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation

Published on: October 4, 2024

723
Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons
08:20

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

Published on: March 28, 2016

8.0K

Related Experiment Videos

Last Updated: Sep 27, 2025

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
09:32

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development

Published on: June 15, 2017

8.8K
Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation
08:00

Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation

Published on: October 4, 2024

723
Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons
08:20

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

Published on: March 28, 2016

8.0K

Area of Science:

  • Cellular electrophysiology
  • Molecular biology
  • Ion channel function

Background:

  • Epithelial sodium channel (ENaC) activity is regulated by the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2).
  • ENaC contains two putative PIP2 binding sites, βN1 and γN2, located in the N termini of its β and γ subunits.
  • The functional significance of these PIP2 binding sites in a cellular context remains unclear.

Purpose of the Study:

  • To investigate the role of the βN1 and γN2 PIP2 binding sites in regulating ENaC activity in response to cellular PIP2 level changes.
  • To compare the effects of PIP2 depletion and recovery on wild-type and mutant ENaC activity and intracellular sodium levels ([Na+]i).

Main Methods:

  • Utilized an optogenetic system (CIBN/CRY2-OCRL) for inducible depletion and recovery of PIP2.
  • Performed whole-cell patch clamp electrophysiology to measure ENaC channel activity.
  • Monitored intracellular sodium levels ([Na+]i) using the fluorescent indicator CoroNa Green AM.

Main Results:

  • Mutations in either the βN1 or γN2 PIP2 binding sites, or both, rendered ENaC unresponsive to PIP2 depletion and recovery, as assessed by electrophysiology.
  • While βN1 mutants showed no change in CoroNa Green fluorescence, γN2 mutants exhibited altered [Na+]i readings, attributed to the dye's uptake and half-life.
  • Electrophysiology confirmed that both βN1 and γN2 sites are individually essential for maximal ENaC activity when PIP2 is present.

Conclusions:

  • The βN1 and γN2 sites are critical for mediating PIP2's regulatory effects on ENaC channel activity.
  • Results highlight the necessity of these specific PIP2 binding sites for proper ENaC function in a cellular environment.
  • Caution is advised when interpreting intracellular sodium measurements using CoroNa Green AM due to potential variations in dye kinetics.