Action of rat liver Gal beta 1-4GlcNAc alpha(2-6)-sialyltransferase on Man beta 1-4GlcNAc beta-OMe, GalNAc beta 1-4GlcNAc beta-OMe, Glc beta 1-4GlcNAc beta-OMe and GlcNAc beta 1-4GlcNAc beta-OMe as synthetic substrates.

Incubation of synthetic Man beta 1-4GlcNAc beta-OMe, GalNAc beta 1-4GlcNAc beta-OMe, Glc beta 1-4GlcNAc beta-OMe, and GlcNAc beta 1-4GlcNAc beta-OMe with CMP-Neu5Ac and rat liver Gal beta 1-4GlcNAc alpha(2-6)-sialyltransferase resulted in the formation of Neu5Ac alpha 2-6Man beta 1-4GlcNAc beta-OMe, Neu5Ac alpha 2-6GalNAc beta 1-4GlcNAc beta-OMe, Neu5Ac alpha 2-6Glc beta 1-4GlcNAc beta-OMe and Neu5Ac alpha 2-6GlcNAc beta 1-4GlcNAc beta-OMe, respectively. Under conditions which led to quantitative conversion of Gal beta 1-4GlcNAc beta-OEt into Neu5Ac alpha 2-6Gal beta 1-4GlcNAc beta-OEt, the aforementioned products were obtained in yields of 4%, 48%, 16% and 8%, respectively. HPLC on Partisil 10 SAX was used to isolate the various sialyltrisaccharides, and identification was carried out using 1- and 2-dimensional 500-MHz 1H-NMR spectroscopy.