Related Experiment Video

Updated: Jan 21, 2026

Optimized Transfection Strategy for Expression and Electrophysiological Recording of Recombinant Voltage-Gated Ion Channels in HEK-293T Cells
12:59

Optimized Transfection Strategy for Expression and Electrophysiological Recording of Recombinant Voltage-Gated Ion Channels in HEK-293T Cells

Published on: January 19, 2011

33.4K

A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary CaVβ subunits

Travis J Morgenstern1, Jinseo Park1, Qing R Fan1

  • 1Department of Pharmacology, Columbia University, Vagelos College of Physicians and Surgeons, New York, United States.

Elife
|August 13, 2019
PubMed

Insights

Researchers developed a novel genetically-encoded inhibitor, CaV-aβlator, targeting high-voltage-activated calcium channels (HVACCs). This innovative tool offers precise, versatile inhibition for cardiovascular and neurological disease therapies.

Area of Science:

  • Molecular Biology
  • Neuroscience
  • Cardiology

Background:

  • High-voltage-activated calcium channels (HVACCs) are crucial drug targets for cardiovascular and neurological conditions.
  • Existing small-molecule inhibitors lack tissue-specificity and versatility.
  • Genetically-encoded inhibitors offer potential for enhanced precision and application range.

Purpose of the Study:

  • To engineer a novel, genetically-encoded inhibitor for high-voltage-activated calcium channels (HVACCs).
  • To assess the efficacy and mechanism of the engineered inhibitor in various cell types and tissues.
  • To establish a generalizable platform for developing genetically-encoded modulators of membrane protein complexes.

Main Methods:

  • Isolation of a llama nanobody (nb.F3) targeting HVACC CaVβ subunits.
  • Fusion of nb.F3 with the Nedd4L E3 ubiquitin ligase HECT domain to create CaV-aβlator.
  • Expression and functional assessment of CaV-aβlator in HEK293 cells, cardiomyocytes, neurons, and pancreatic β cells.
  • Analysis of CaV1.2 channel trafficking in cardiomyocytes using immunofluorescence microscopy.

Main Results:

  • The engineered CaV-aβlator effectively inhibited currents from diverse HVACCs in reconstituted systems and endogenous channels in native cells.
  • CaV-aβlator demonstrated efficacy in cardiomyocytes, neurons, and pancreatic β cells.
  • In cardiomyocytes, CaV-aβlator induced the redistribution of CaV1.2 channels from dyads to late endosomes.

Conclusions:

  • CaV-aβlator represents a potent, genetically-encoded inhibitor of HVACCs.
  • This study presents a versatile platform for engineering genetically-encoded modulators of macro-molecular membrane protein complexes.
  • The findings open new avenues for targeted therapies in cardiovascular and neurological diseases.

Keywords:
Nedd4biochemistrycalcium channelcalcium channel betacavia porcelluscell biologychemical biologymousenanobodyubiquitin

More Related Videos

Voltage and Calcium Dual Channel Optical Mapping of Cultured HL-1 Atrial Myocyte Monolayer08:25

Voltage and Calcium Dual Channel Optical Mapping of Cultured HL-1 Atrial Myocyte Monolayer

Published on: March 23, 2015

10.8K
Profiling Voltage-gated Potassium Channel mRNA Expression in Nigral Neurons using Single-cell RT-PCR Techniques07:31

Profiling Voltage-gated Potassium Channel mRNA Expression in Nigral Neurons using Single-cell RT-PCR Techniques

Published on: September 27, 2011

15.6K

Related Concept Videos

Voltage-gated Ion Channels01:26

Voltage-gated Ion Channels

Voltage-gated ion channels are transmembrane proteins that open and close in response to changes in the membrane potential. They are present on the membranes of all electrically excitable cells such as neurons, heart, and muscle cells.
Generally, all voltage-gated ion channels have a 'voltage-sensing domain' that spans the lipid bilayer. The charged residues in the sensor move in response to the membrane potential changes that open the channel allowing ions movement. There are several types of...
10.5K
Non-gated Ion Channels01:24

Non-gated Ion Channels

Ion channels are specialized proteins on the plasma membrane that allow charged ions to pass down their electrochemical gradient. Their main function is to maintain the membrane potential which is critical for cell viability. These channels are either gated or non-gated and can transport more than a thousand ions within milliseconds for the cellular event to occur.
Compared to the gated ion channels, the non-gated channels, also known as leakage or passive channels, have no gating mechanism....
8.0K
Ligand-gated Ion Channels01:19

Ligand-gated Ion Channels

Ligand-gated ion channels are transmembrane proteins with a channel for ions to pass through and a binding site for a ligand. The channel opens only when a ligand attaches to the binding site.
Three Subfamilies of Ligand-gated Ion Channels
Ligand-gated ion channels fall into three subfamilies. The 'Cys-loop' includes the nicotinic acetylcholine receptors, γ-aminobutyric acid (GABA), glycine, and 5-hydroxytryptamine receptors. The second one is the 'Pore-loop' channels that...
14.0K
Mechanically-gated Ion Channels01:12

Mechanically-gated Ion Channels

Mechanically-gated ion channels are proteins found in eukaryotic and prokaryotic cell membranes that open in response to mechanical stress. Tension, compression, swelling, and shear stress can alter the conformation of the protein, opening a transmembrane channel that allows the passage of ions for signal transmission. In eukaryotes, mechanically-gated channels are distributed in several regions like the neurons, lungs, skin, bladder, and heart, where they play critical roles in numerous...
7.6K
Ligand-Gated Ion Channel Receptor: Gating Mechanism01:30

Ligand-Gated Ion Channel Receptor: Gating Mechanism

Ligand-gated ion channels are transmembrane proteins that play a vital role in intercellular communication and functions of the nervous system. They allow the influx of ions across the membrane once the neurotransmitter binds, allowing the subsequent transmission of electrical excitation across the neurons. Other ligand-gated ion channels, like the γ-aminobutyric acid (GABA) receptor, permit anions like chloride into the cells on the binding of the GABA molecule. Their entry into the cell...
3.8K
G-Protein Gated Ion Channels01:21

G-Protein Gated Ion Channels

GPCRs are primarily responsible for our sense of smell, taste, and vision.  The binding of a sensory stimulus activates GPCR to stimulate effector proteins, many of which are ion channels in the sensory organs. GPCRs modulate the opening and closing of the target ion channels either directly by binding them, or by releasing second messengers that activate these channels. As ions move across the membrane, the membrane potential is altered, which induces an appropriate response.
Sensory...
5.6K