Related Experiment Video
Updated: Aug 28, 2026

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
Published on: October 4, 2017
Endogenous glycosyltransferases glucosylate lipids in flagella of Euglena
Abstract:
Flagella, intact deflagellated cells and isolated cell surfaces of the unicell , Euglena were separately assayed for glycosyltransferase activity by incubating these fractions with uridine diphosphate-[3H]glucose and isolating radiolabeled products. Most of the label was incorporated into lipophilic products, soluble in chloroform/methanol, which could be separated via thin layer chromatography or LH-60 chromatography into four distinct classes. The most polar of these products was extracted from flagella and purified by column chromatography for use as an in vitro substrate to identify flagella-associated glycosyltransferases. After flagella were treated with the detergent CHAPS , a soluble fraction was removed that was capable of glycosylation in solution. The glycosyltransferase(s) responsible for this activity were further enriched on sucrose or fructose gradients and ultimately identified on acrylamide gels through the combined use of nondenaturing gels, dial-[3H]uridine diphosphate binding, and fluorography. The enzyme had an apparent monomer molecular weight of 32,000 and consisted of four or fewer subunits. The occurrence of endogenous glycosyltransferase(s) in flagella suggests that modifications and/or assembly of the flagella surface can take place in situ in this organism.
Related Concept Videos
Lipids as Anchors
The carboxy-terminal of most of the prenylated proteins, such as Ras proteins, contains the...
Protein Glycosylation
Glycosylation occurs in...
Oligosaccharide Assembly
Multiple sugar molecules that may or may...
Protein Folding Quality Check in the RER
GPI Anchoring of Proteins in the ER Membrane
GPI-anchor structure
A sequence of 11 enzymatic reactions results in the synthesis of the complete GPI anchor consisting of a hydrophobic and a hydrophilic portion. The hydrophobic portion comprises phosphatidylinositol, while the hydrophilic part comprises polar groups like phosphoethanolamine,...
Proteoglycans

