Wheat germ agglutinin chromatography of GlcNacβ1-3(GlcNAcβl-6)Gal and GlcNAcβ1-3(GlcNAcβ1-6)Galβ1-4GlcNAc, obtained by in vitro synthesis and by partial cleavage of teratocarcinoma poly-N-acetyllactosaminoglycans

Abstract

GlcNAcβ1-3(GlcNAcβ1-6)[14C(U)]Gal and GlcNAcβ1-3(GlcNAcβ1-6)[14C(U)]Galβ1-4GlcNAc were prepared by in vitro synthesis. They were characterized by enzymatic sequencing, by partial acid hydrolysis, and by periodate oxidation experiments. The two saccharides were isolated also from partial acid hydrolysates of metabolically labeled poly-N- acetyllactosaminoglycans of murine embryonal carcinoma cells (line PC 13). The tetrasaccharide was retarded in a column of agarose-linked wheat germ agglutinin; the trisaccharide was strongly bound. Chromatography in this column separated the trisaccharide into two distinct peaks, which represented interconvertible molecules. Together with our previous data on linear teratocarcinoma saccharides, these findings show that affinity chromatography with immobilized wheat germ agglutinin can be advantageously used in fractionating radiolabeled oligo-N-acetyllactosaminoglycans and saccharides related to them.Key words: GlcNAcβ1-3(GlcNAcβ1-6)Gal, GlcNAcβ1-3(GlcNAcβ1-6)Galβ1-4GlcNAc, wheat germ agglutinin – agarose chromatography, in vitro biosynthesis, teratocarcinoma cell.