Bi-antennary oligo-(N-acetyllactosamino)glycans of I-type are galactosylated preferentially at the GlcNAc beta 1-6Gal linked arms by alpha 1,3-galactosyltransferase of bovine thymus.
alpha 1,3-Galactosylation of radiolabelled bi-antennary acceptors Gal beta 1-4GlcNAc beta 1-3(Gal beta 1-4GlcNAc beta 1-6)Gal-R (R = 1-OH, beta 1-4GlcNAc or beta 1-4Glc) with bovine thymus alpha 1,3-galactosyltransferase was studied. At all stages of the reactions the three acceptors reacted faster at the 1-->6 linked arm than at the 1-->3 linked branch. Hence, in addition to the doubly alpha 1,3-galactosylated products, practically pure Gal beta 1-4GlcNAc beta 1-3(Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6)Gal-R could be obtained from the three acceptors in reactions that had proceeded to near completion. The isomeric mono-alpha 1,3-galactosylated products were identified by using exoglycosidases to remove the branches unprotected by alpha 1,3-galactoses and by subsequently identifying the resulting linear glycans chromatographically.