Figure 1
Analysis of perlecan GAG-composition with SDS-PAGE.
(a) Schematic illustration of perlecan. The proteoglycan may be substituted with GAGs at various positions, including three HS-chains at the N-terminal end as well as a CS-chain at the C-terminal end18. The number of disaccharides (n) and the degree of sulfation may vary at each GAG-position. (b) Trypsin-digested mouse perlecan was applied onto a SAX-column and enriched for GAG-glycopeptides (see Methods). The GAG-glycopeptides were eluted from the column with increasing salt concentrations (0.4 M, 0. 8 M and 1.6 M NaCl) and analyzed with SDS-PAGE / Alcian blue. Perlecan incubated with and without trypsin was loaded as control. (c,d) An identical set of SAX-enriched GAG-glycopeptides were treated with either chondroitinase ABC (CSase) (c) or heparinase (HSase) (d) prior to the SDS-PAGE / Alcian blue analysis.
