Related Experiment Video

Updated: Aug 24, 2026

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
08:27

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

Published on: January 7, 2019

Control of TRPC and store-operated channels by protein kinase C

Kartik Venkatachalam1, Fei Zheng, Donald L Gill

  • 1Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 108 North Greene Street, Baltimore, MD 21201, USA.

Novartis Foundation Symposium
|April 24, 2004
PubMed

Insights

Protein kinase C (PKC) activation by diacylglycerol (DAG) inhibits TRPC5 and TRPC4 channels, but stimulates TRPC3 channels. This distinct regulation by PKC and DAG influences TRPC channel activity and calcium signaling.

Area of Science:

  • Molecular Biology
  • Cell Physiology
  • Biochemistry

Background:

  • Transient Receptor Potential (TRPC) channels are crucial for calcium (Ca2+) signaling.
  • TRPC channel function is modulated by various cellular mechanisms.

Purpose of the Study:

  • To investigate the role of protein kinase C (PKC) and diacylglycerol (DAG) in regulating TRPC channel activity.
  • To elucidate the distinct mechanisms by which DAG and PKC control different TRPC subtypes.

Main Methods:

  • Utilized HEK293 and DT40 cell lines.
  • Manipulated diacylglycerol (DAG) levels through pharmacological inhibitors and activators.
  • Assessed TRPC channel activation and deactivation using electrophysiological techniques.
  • Investigated the involvement of PKC using specific inhibitors and activators.

Main Results:

  • DAG-induced PKC activation inhibited TRPC5 and TRPC4 channel activation in a PKC-dependent manner.
  • DAG stimulated TRPC3 channel activity independently of PKC.
  • PKC activation blocked TRPC3 channel activation, while PKC inhibition decreased TRPC3 deactivation.
  • Store-operated calcium entry was reduced by DAG or DAG-lipase inhibition in a PKC-dependent manner.

Conclusions:

  • DAG-induced PKC activation represents a universal feedback mechanism inhibiting TRPC channels.
  • Distinct regulatory pathways involving PKC and DAG control the activation of different TRPC subtypes.
  • These regulatory mechanisms contribute to the diverse functional phenotypes of TRPC channels in cellular signaling.

More Related Videos

Purification of Endogenous Drosophila Transient Receptor Potential Channels08:39

Purification of Endogenous Drosophila Transient Receptor Potential Channels

Published on: December 28, 2021

Controllable Ion Channel Expression through Inducible Transient Transfection10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

Related Concept Videos

Calmodulin-dependent Signaling01:16

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...
GPCRs Regulate Adenylyl Cylase Activity01:09

GPCRs Regulate Adenylyl Cylase Activity

Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of cells.
Two...
IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and produces two-second...
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...