Related Experiment Video

Updated: Dec 6, 2025

Yeast Luminometric and Xenopus Oocyte Electrophysiological Examinations of the Molecular Mechanosensitivity of TRPV4
12:09

Yeast Luminometric and Xenopus Oocyte Electrophysiological Examinations of the Molecular Mechanosensitivity of TRPV4

Published on: December 31, 2013

10.5K

TRIM4 interacts with TRPM8 and regulates its channel function through K423-mediated ubiquitination

Yuan Huang1, Shunyao Li1, Zhenhua Jia1

  • 1National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan, China.

Journal of Cellular Physiology
|October 10, 2020
PubMed

Insights

Tripartite motif-containing 4 (TRIM4) downregulates the TRPM8 channel by promoting its ubiquitination. This process requires UBA1 and targets a specific lysine residue, affecting TRPM8 function and cell surface expression.

Area of Science:

  • Molecular Biology
  • Cell Physiology
  • Ion Channel Function

Background:

  • Transient receptor potential melastatin member 8 (TRPM8) is a cold-activated cation channel involved in pain signaling (allodynia).
  • TRPM8 dysfunction and abnormal expression are implicated in human cancers.
  • The regulatory mechanisms of TRPM8, particularly ubiquitination, are not well understood.

Purpose of the Study:

  • To investigate the role of ubiquitination in regulating TRPM8 channel function.
  • To identify novel interaction partners of TRPM8 involved in its ubiquitination and regulation.
  • To elucidate the specific molecular mechanisms by which TRIM4 affects TRPM8.

Main Methods:

  • Co-immunoprecipitation to identify TRIM4 as a TRPM8 interaction partner.
  • Patch-clamp electrophysiology to assess TRPM8 channel activity.
  • Western blotting and immunofluorescence to analyze TRPM8 expression and ubiquitination.
  • Site-directed mutagenesis to identify the ubiquitination site.

Main Results:

  • TRIM4 directly interacts with TRPM8 via its SPRY domain.
  • TRIM4 negatively regulates TRPM8 currents and reduces cell surface expression.
  • TRIM4 promotes K63-linked ubiquitination of TRPM8 at lysine residue 423.
  • UBA1 (Ub-activating enzyme E1) interacts with TRPM8 and is required for TRIM4-mediated regulation.

Conclusions:

  • TRIM4 acts as an E3 ubiquitin ligase that downregulates TRPM8 channel activity and surface expression.
  • TRIM4-mediated TRPM8 regulation occurs through K423 ubiquitination and requires UBA1.
  • This study reveals a novel ubiquitination-dependent pathway for TRPM8 regulation.
Keywords:
TRIM4TRPM8UBA1patch clampubiquitination

More Related Videos

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice08:35

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Published on: March 17, 2015

15.4K
Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy08:27

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

9.8K

Related Concept Videos

Thermosensation01:43

Thermosensation

Peripheral thermosensation is the perception of external temperature. A change in temperature (on the surface of the skin and other tissues) is detected by a family of temperature-sensitive ion channels called Transient Receptor Potential, or TRP, receptors. These receptors are located on free nerve endings. Those detecting cold temperatures are closer to the surface of the skin than the nerve endings detecting warmth. These thermoTRP channels, while temperature selective, have relatively...
33.3K
Enzyme-linked Receptors01:00

Enzyme-linked Receptors

Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...
84.2K
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...
7.4K
Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
3.5K
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
8.3K
Receptor Tyrosine Kinases01:26

Receptor Tyrosine Kinases

Receptor tyrosine kinases or RTKs are membrane-bound receptors that phosphorylate specific tyrosine on protein substrates. RTKs regulate cellular growth, differentiation, survival, and migration. They contain an extracellular ligand binding domain, a transmembrane domain, and a cytosolic tail with intrinsic kinase activity. Several extracellular signaling molecules activate RTKs in one or more ways and relay the signal downstream. Ligands such as platelet-derived growth factor (PDGF) or...
16.9K