Fig. 3: In vitro chain elongation assay using purified CS polymerase complexes.
From: Structural basis for human chondroitin sulfate chain polymerization
a Schematic illustration of an in vitro glycosylation assay used to monitor chain elongation by the CS polymerase complexes. The fluorescently labelled Penta-CSF1 peptide, derived from the CSPG colony-stimulating factor 1, served as the acceptor substrate, while UDP-GlcA and UDP-GalNAc were used as donor substrates. Monosaccharide symbols follow the SNFG (Symbol Nomenclature for Glycans) system67. b Catalytic activity of purified CS polymerase complexes was studied using the in vitro glycosylation assay described in (a). Reaction products were separated by polyacrylamide gel electrophoresis, and fluorescent peptides were visualized using a fluorescence imager. This result is representative of three independent experiments. c Analysis of CHSY3-CHPF reaction product upon chondroitinase ABC treatment from a single experiment. Tetra-, penta-, hexa-, and heptasaccharide peptides were loaded alongside as length markers. Source data are provided as a Source Data file.
