Related Experiment Video

Updated: May 12, 2026

Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark
10:09

Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark

Published on: January 26, 2018

7.4K

Evidence for Functional Regulation of the KLHL3/WNK Pathway by O-GlcNAcylation

Jimin Hu1, Duc T Huynh1, Denise E Dunn2

  • 1Department of Biochemistry, Duke University School of Medicine, Durham, North Carolina, USA.

Biorxiv : the Preprint Server for Biology
|March 10, 2025
PubMed

Insights

O-linked N-acetylglucosamine (O-GlcNAc) modifies Kelch-like (KLHL) proteins, including KLHL3, and WNK kinases. This glycosylation impacts WNK4 function, affecting osmolarity control and ferroptosis, crucial for health.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Signaling

Background:

  • Kelch-like (KLHL) proteins are crucial adaptors for E3 ubiquitin ligase complexes, regulating substrate stability and proteostasis.
  • Dysregulation of KLHL proteins is implicated in diseases like cancer, neurodegeneration, and hypertension.
  • Previous studies indicated O-linked N-acetylglucosamine (O-GlcNAc) modifies KEAP1 and gigaxonin, suggesting a broader role for this post-translational modification in KLHL regulation.

Purpose of the Study:

  • To investigate the role of O-GlcNAc in the regulation of the KLHL3/WNK kinase axis.
  • To determine if KLHL3 and WNK kinases are O-GlcNAcylated.
  • To explore the functional consequences of O-GlcNAc modification on WNK4 activity.

Main Methods:

  • Biochemical assays to assess protein interactions and modifications.
  • Glycoproteomic analysis to identify O-GlcNAcylated proteins.
  • Functional assays to evaluate the impact of O-GlcNAc on WNK4-mediated processes.

Main Results:

  • Biochemical and glycoproteomic data confirmed O-GlcNAcylation of human KLHL3 and WNK1-4 kinases.
  • O-GlcNAcylation was found to influence WNK4 function in regulating osmolarity control.
  • Evidence suggests O-GlcNAc modification of the KLHL3/WNK axis impacts ferroptosis, with implications for blood pressure and neuronal health.

Conclusions:

  • O-GlcNAcylation functionally regulates the KLHL3/WNK signaling pathway.
  • This study supports a model where O-GlcNAc broadly regulates KLHL proteins and proteostasis.
  • The findings highlight potential therapeutic targets for diseases involving KLHL and WNK pathway dysregulation.

More Related Videos

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins08:12

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

Published on: January 8, 2018

11.2K
Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting10:08

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Published on: December 9, 2022

2.1K

Related Concept Videos

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.
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
Oligosaccharide Assembly01:24

Oligosaccharide Assembly

Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
Canonical Wnt Signaling Pathway02:54

Canonical Wnt Signaling Pathway

The gene encoding the main signaling molecules of the Wnt signaling pathways (the Wnt proteins) was discovered almost four decades ago by Nüsslein-Volhard and Wieschaus. They identified and originally named the gene "wingless" (wg) after a phenotype discovered during their landmark genetic screen in Drosophila for body pattern defects. At around the same time, another researcher named Harold Varmus found that a murine tumor virus activates the mammalian wg homolog, Int-1, which results in tumor...
Non-Canonical Wnt Signaling Pathways01:41

Non-Canonical Wnt Signaling Pathways

Wnt is a zygotic effect gene that is expressed during very early embryonic development. It regulates various processes in animals starting from early development through the adult stage, such as organogenesis in the embryo and maintenance of neuronal and blood stem cells. Wnt proteins can induce a wide variety of intracellular pathways depending upon the specific abilities of different Wnt ligands to form a complex with shared and cognate receptors in the presence of different co-receptors. The...
Proteoglycans01:05

Proteoglycans

Glycans, a class of complex heterogeneous molecules, can be covalently attached to proteins to form glycosylated proteins that regulate various physiological and pathological processes. Glycosylated proteins or glycoproteins comprise N-linked and O-linked oligosaccharides. O-glycosylation is the most common type of protein glycosylation. Here, glycans attach to the oxygen atom of the hydroxyl groups of Serine or Threonine residues. O-linked glycosylation occurs later in protein processing,...