Related Experiment Video

Updated: Aug 9, 2026

Inducible and Reversible Dominant-negative DN Protein Inhibition
08:35

Inducible and Reversible Dominant-negative DN Protein Inhibition

Published on: January 7, 2019

8.8K

CRAC channel is inhibited by neomycin in a Ptdlns(4,5)P2-independent manner

Kun Huang1, Xuemei Wang, Yanjun Liu

  • 1Department of Ultrasound, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning, China.

Cell Biochemistry and Function
|February 10, 2015
PubMed

Insights

Neomycin inhibits store-operated calcium entry via CRAC channels. This inhibition is independent of neomycin

Area of Science:

  • Cellular biology
  • Molecular pharmacology
  • Ion channel physiology

Background:

  • Store-operated calcium entry (SOCE) is crucial for cellular calcium homeostasis.
  • Calcium release-activated calcium (CRAC) channels mediate SOCE.
  • Neomycin, an aminoglycoside, is known to bind phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2).

Purpose of the Study:

  • To investigate the mechanism by which neomycin inhibits CRAC channel activity.
  • To determine the role of PtdIns(4,5)P2 in neomycin-mediated inhibition of CRAC channels.

Main Methods:

  • Patch clamp electrophysiology to record CRAC currents.
  • Co-expression of Orai1 and Stim1 in HEK293 cells to reconstitute CRAC channels.
  • Utilizing a rapamycin-inducible PtdIns(4,5)P2-specific phosphatase (Inp54p) system to manipulate plasma membrane PtdIns(4,5)P2 levels.

Main Results:

  • Neomycin inhibited CRAC currents reconstituted by Orai1 and Stim1.
  • Depletion of PtdIns(4,5)P2 using the Inp54p system did not alter CRAC channel activity.
  • Neomycin's inhibition of CRAC channels was independent of its binding to PtdIns(4,5)P2.

Conclusions:

  • Phosphatidylinositol 4,5-bisphosphate is not required for CRAC channel function.
  • Neomycin inhibits CRAC channels through a mechanism independent of PtdIns(4,5)P2 binding.
Keywords:
CRAC channelCa2+Orai1PtdIns(4,5)P2Stim1neomycin

More Related Videos

Native Cell Membrane Nanoparticles System for Membrane Protein-Protein Interaction Analysis07:31

Native Cell Membrane Nanoparticles System for Membrane Protein-Protein Interaction Analysis

Published on: July 16, 2020

6.7K
Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time08:33

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

Published on: March 11, 2021

2.4K

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...