Related Experiment Video
Updated: May 15, 2026
20:23
A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Published on: October 1, 2009
Tandem lectin weak affinity chromatography for glycoprotein enrichment
Zhi Yuan Ma1, Yuliya Skorobogatko, Keith Vosseller
1Department of Biochemistry and Molecular Biology, Drexel University, Philadelphia, PA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 9, 2013
Summary
Lectin weak affinity chromatography (LWAC) enriches O-GlcNAc modified peptides. This method aids in identifying challenging O-GlcNAc attachment sites on intracellular proteins using mass spectrometry.
Area of Science:
- Biochemistry
- Proteomics
- Post-translational modifications
Background:
- O-linked β-N-acetylglucosamine (O-GlcNAc) is a crucial intracellular post-translational modification.
- The low stoichiometry of O-GlcNAc modification presents challenges in identifying modification sites.
- Understanding O-GlcNAc dynamics is vital for cellular signaling and disease research.
Purpose of the Study:
- To describe the application of lectin weak affinity chromatography (LWAC) for enriching O-GlcNAc modified peptides.
- To provide a method for mapping O-GlcNAc attachment sites on proteins.
- To facilitate the study of O-GlcNAc modification in biological systems.
Main Methods:
- Digestion of protein samples with trypsin.
- Application of lectin weak affinity chromatography (LWAC) using wheat germ agglutinin.
- Enrichment of O-GlcNAc-modified peptides.
- Identification of enriched peptides using mass spectrometry.
Main Results:
- LWAC effectively enriches peptides containing O-GlcNAc modifications.
- The method allows for the isolation of low-stoichiometry modified peptides.
- Enriched peptides are suitable for subsequent mass spectrometry analysis.
Conclusions:
- LWAC is a powerful technique for the enrichment and identification of O-GlcNAc modified peptides.
- This approach simplifies the challenging task of mapping O-GlcNAc sites.
- The method has significant implications for studying the biological roles of O-GlcNAcylation.
Related Concept Videos
01:03Affinity Chromatography
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
01:33Peptide Identification Using Tandem Mass Spectrometry
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...