Related Experiment Video
Updated: Mar 27, 2026
11:39
Identifying Cell Surface Markers of Primary Neural Stem and Progenitor Cells by Metabolic Labeling of Sialoglycan
Published on: September 7, 2019
5.8K
Selective Exo-Enzymatic Labeling Detects Increased Cell Surface Sialoglycoprotein Expression upon Megakaryocytic
Seok-Ho Yu1, Peng Zhao1, Tiantian Sun1
1From the Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602.
The Journal of Biological Chemistry
|January 7, 2016
Summary
Selective exo-enzymatic labeling (SEEL) efficiently labels cell surface glycans, improving glycoprotein detection. This method revealed distinct glycoprotein profiles and increased sialylated cell adhesion molecules during human erythroleukemia cell differentiation.
Area of Science:
- Glycobiology
- Proteomics
- Cell Biology
Background:
- Metabolic labeling of cell surface glycans can suffer from low precursor incorporation.
- Selective exo-enzymatic labeling (SEEL) offers a specific alternative using glycosyltransferases and nucleotide-sugar analogs.
- Understanding cell surface glycan changes during differentiation is crucial for cell biology.
Purpose of the Study:
- To compare SEEL efficiency for N- and O-glycan sialylation in undifferentiated and differentiated human erythroleukemia cells (HEL).
- To investigate changes in sialoglycoprotein profiles during HEL cell differentiation into megakaryocytes.
- To assess the utility of SEEL in identifying cell surface glycoproteins and adhesion molecules.
Main Methods:
- Selective exo-enzymatic labeling (SEEL) using sialyltransferases ST6Gal1 (N-glycans) and ST3Gal1 (O-glycans).
- Neuraminidase treatment to enhance SEEL efficiency by increasing acceptor sites.
- Glycoprotein enrichment via immunoprecipitation followed by mass spectrometry identification.
- Comparison of glycoprotein profiles between undifferentiated and differentiated HEL cells.
Main Results:
- SEEL efficiency was significantly enhanced by prior neuraminidase treatment.
- Differentiation of HEL cells to megakaryocytes resulted in a shift towards N-linked sialoglycoproteins.
- SEEL improved the detection of many glycoproteins and allowed identification of unique species using specific sialyltransferases.
- Increased surface expression of sialylated cell adhesion molecules, including integrinβ3 and CD44, was observed upon differentiation.
Conclusions:
- SEEL is an effective method for labeling cell surface glycans, overcoming limitations of metabolic labeling.
- SEEL facilitates the identification of distinct glycoprotein profiles associated with cellular differentiation.
- The study identified specific changes in surface sialylated glycoproteins, including key adhesion molecules, during megakaryocyte differentiation.
