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Oligosaccharide Assembly

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...

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Related Experiment Video

Updated: Apr 30, 2026

Improved In-gel Reductive β-Elimination for Comprehensive O-linked and Sulfo-glycomics by Mass Spectrometry

Improved In-gel Reductive β-Elimination for Comprehensive O-linked and Sulfo-glycomics by Mass Spectrometry

Published on: November 20, 2014

Detecting O-GlcNAc using in vitro sulfation.

Zhengliang L Wu¹, Matthew T Robey², Timothy Tatge²

¹R&D Systems, Inc., 614 McKinley Place N.E., Minneapolis, MN 55413, USA leon.wu@rndsystems.com jzaia@bu.edu.

|May 7, 2014

Summary

This summary is machine-generated.

Detecting O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation is challenging. A new method uses in vitro sulfation with specific enzymes and radioisotopes for sensitive and convenient O-GlcNAc detection.

Keywords:

CHST O-GlcNAc OGA OGT glycosylation

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Last Updated: Apr 30, 2026

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Detection of Protein S-Acylation using Acyl-Resin Assisted Capture

Detection of Protein S-Acylation using Acyl-Resin Assisted Capture

Published on: April 10, 2020

Area of Science:

Biochemistry
Molecular Biology
Glycobiology

Background:

O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation is a crucial post-translational modification involved in vital cellular processes and diseases.
Detecting O-GlcNAc modifications on proteins presents significant technical challenges in biological research.

Purpose of the Study:

To develop and validate a novel, sensitive, and convenient method for detecting O-GlcNAc glycosylation.
To utilize in vitro sulfation with specific sulfotransferases and a radioisotope for enhanced O-GlcNAc detection.

Main Methods:

Employing N-acetylglucosamine (GlcNAc)-specific sulfotransferases (CHST2 and CHST4) for in vitro sulfation of O-GlcNAc residues.
Utilizing the radioisotope (35)S to label sulfated O-GlcNAc on free GlcNAc, peptides, and nuclear/cytoplasmic proteins.
Analyzing sulfation sensitivity to O-GlcNAc transferase (OGT) and O-β-N-acetylglucosaminidase, followed by SDS-PAGE and autoradiography.

Main Results:

Successful demonstration of sulfation on free GlcNAc and O-GlcNAc residues of various proteins.
Confirmation that the sulfation process is sensitive to enzymatic treatments by OGT and O-β-N-acetylglucosaminidase.
Validation of the method's sensitivity, specificity, and convenience through gel electrophoresis and autoradiography.

Conclusions:

The described in vitro sulfation method provides a sensitive, specific, and convenient approach for O-GlcNAc detection.
This method offers a valuable tool for studying the biological roles and disease implications of O-GlcNAc glycosylation.