Histone Serine/Threonine O-GlcNAcylation Analysis

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Histone O-GlcNAcylation is a vital nutrient-sensitive modification that directly integrates cellular metabolic status with chromatin regulation. CD BioSciences offers specialized histone serine/threonine O-GlcNAcylation analysis services to help you uncover the functional roles of this dynamic modification in metabolic signaling, disease mechanisms, and cellular adaptation. Our tailored solutions provide precise, quantitative mapping of O-GlcNAc sites to reveal how metabolic flux rewires the epigenetic landscape in health and disease.

Introduction to Histone Serine/Threonine O-GlcNAcylation

Histone O-GlcNAcylation is a reversible, monosaccharide post-translational modification involving the attachment of a single N-acetylglucosamine (O-GlcNAc) molecule to specific serine or threonine residues on histone proteins. This modification is dynamically regulated by two opposing enzymes: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Serving as a key nutrient sensor, its levels are directly influenced by the flux through the hexosamine biosynthetic pathway (HBP), which integrates inputs from glucose, amino acid, fatty acid, and nucleotide metabolism. Functionally, O-GlcNAcylation plays crucial roles in fine-tuning gene expression, modulating the cell cycle, and coordinating cellular stress responses. It often engages in crosstalk or competitive interplay with phosphorylation at the same or adjacent sites, adding a complex layer to the "histone code." Dysregulation of this modification is strongly implicated in the pathogenesis of diabetes, cancer, neurodegenerative diseases, and cardiovascular disorders, highlighting its significance at the intersection of metabolism and epigenetics.

O-GlcNAc glycosylation participates in the regulation of gene expression through different mechanisms.

Fig.1 O-GlcNAcylation is involved in the regulation of gene expression through different mechanisms. (Dupas T, et al., 2023)

Our Services

With a dedicated focus on the intricate interface of metabolism and epigenetics, CD BioSciences provides comprehensive histone serine/threonine O-GlcNAcylation analysis services by integrating high-specificity chemoenzymatic enrichment with advanced mass spectrometry platforms featuring electron transfer dissociation (ETD). Our services are designed to overcome the challenges of detecting this low-stoichiometry and labile modification, delivering reliable data to explore its regulation and function in health and disease.

Customized Solution for Histone Serine/Threonine O-GlcNAcylation Analysis

Our tailored solutions are designed to address the specific challenges of studying the dynamic and metabolism-sensitive histone O-GlcNAcylation. We offer flexible, project-specific strategies that combine optimized enrichment, advanced fragmentation techniques, and robust quantification to deliver precise insights into how nutrient signaling regulates the epigenetic landscape.

Platform Technology Matrix

Technology Tier	Core Technology	Pain Point Addressed
Enrichment Tier	Click Chemistry-based Enrichment	High-specificity capture of O-GlcNAcylated peptides, overcoming low stoichiometry.
Fragmentation Tier	Electron Transfer Dissociation (ETD)	Preserves the intact O-GlcNAc moiety during MS/MS for accurate site localization.
Quantification Tier	Dimethyl Labeling / TMT Quantification	Enables precise measurement of dynamic changes in response to metabolic perturbations.
Validation Tier	Enzymatic Validation (OGA Treatment)	Confirms modification specificity by verifying signal loss upon O-GlcNAcase digestion.

Specialized Service Modules

Global O-GlcNAcome Profiling: Comprehensive, unbiased mapping of O‑GlcNAcylation sites across all histone variants.
Site-Specific Quantification: Targeted absolute or relative quantification of defined O‑GlcNAc sites with high precision.
Metabolic Perturbation Dynamics Analysis: Monitoring O‑GlcNAcylation changes in response to nutrient variations or metabolic inhibitors.
Phosphorylation‑Glycosylation Competition Analysis: Deciphering the interplay between O‑GlcNAcylation and phosphorylation at shared or adjacent residues.
Crosstalk Analysis: Integrated profiling of O‑GlcNAcylation alongside other PTMs to uncover cooperative regulatory networks.
Disease Mechanism Analysis: Comparative O‑GlcNAcome profiling in disease models to identify pathology‑associated modification signatures.

Workflow of Histone Serine/Threonine O-GlcNAcylation Analysis Service

Supported Sample Types

Our platform supports a wide array of sample types, with particular expertise in processing metabolism-relevant and translational research specimens for O‑GlcNAcylation analysis.

Sample Types	Description
Cultured Cells	Mammalian cell lines (e.g., HEK293, HeLa, cancer cell lines) grown under defined metabolic conditions (specific glucose, glutamine, or serum levels). Primary cells (e.g., hepatocytes, neurons, immune cells) under controlled nutrient or inhibitor treatment.
Tissue Specimens	Animal tissues from nutritional or disease models, including liver, pancreas, brain, adipose tissue, and heart. Fresh/frozen clinical tissue biopsies from metabolic or oncology studies (e.g., tumor adjacent normal vs. tumor tissue).
Processed Samples	Pre-extracted and purified histone proteins from any biological source. Chemically or enzymatically modified histone samples (e.g., in vitro O-GlcNAcylated histones for method validation or control experiments).
Archival & Special Samples	Formalin-fixed paraffin-embedded (FFPE) tissue sections (subject to a prior feasibility assessment). Other specimen types (e.g., sorted cell populations, limited clinical samples) are accepted upon consultation and protocol optimization.

*For optimal results, samples should be snap-frozen immediately and shipped on dry ice. We strongly recommend contacting our technical team before submission to confirm specific requirements, minimum amounts, and the most suitable preservation protocol for your research model.

Our Advantages

ETD Mass Spectrometry Expertise: Use of electron transfer dissociation to maintain the O-GlcNAc modification intact during fragmentation, enabling confident site localization—a critical advantage over conventional methods.
High-Efficiency Enrichment Strategies: Optimized chemoenzymatic and immunoaffinity protocols for the sensitive capture of low-abundance O-GlcNAcylated histones.
Metabolic Context Specialization: Deep experience in designing studies that link O-GlcNAcylation changes to alterations in nutrient sensing and metabolic pathways.
End-to-End Scientific Support: Guidance from experimental design through data interpretation by specialists in metabolism and epigenetics.

To further support your exploration of the complex epigenetic landscape, CD BioSciences offers an extensive portfolio of PTM analysis services beyond histone O-GlcNAcylation. We provide solutions for acetylation, methylation, phosphorylation, ubiquitination, SUMOylation, formylation, succinylation, citrullination, crotonylation, and lactylation, among others. Our flexible platform enables the creation of fully customized, multi-dimensional PTM profiling studies tailored to your specific research questions in metabolism, signaling, and disease biology. Contact us to design an integrated analytical strategy for your next project.

Reference

1. Dupas T, Lauzier B, McGraw S. O-GlcNAcylation: the sweet side of epigenetics[J]. Epigenetics & Chromatin, 2023, 16(1): 49.