The widespread effect of beta 1,4-galactosyltransferase on N-glycan processing

We investigated beta1,4-GalT (UDP-galactose: beta -D-N-acetylglucosaminide beta1,4-galactosyltransferase) in terms of intracellular competition with G nT-IV (UDP-N-acetylglucosamine: alpha1,3-D-mannoside beta1,4-N-acetylglucos aminyltransferase) and GnT-V (UDP-N-acetylglucosamine: alpha1,6-D-mannoside beta1,6-N-acetylglucosaminyltransferase). The beta1,4-GalT-I gene was intr oduced into Chinese hamster ovary (CHO) cells producing human interferon (h IFN)-gamma (IM4/V/IV cells) and five clones expressing various levels of be ta1,4-GalT were isolated. As we previously reported, parental IM4/V/IV cell s express high levels of GnT-IVa and -V and produce hIFN-gamma having prima rily tetraantennary sugar chains. The branching of sugar chains on hIFN-gam ma was suppressed in the beta1,4-GalT-enhanced clones to a level correspond ing to the intracellular activity of beta1,4-GalT relative to GnTs. Moreove r, the contents of hybrid-type and high-mannose-type sugar chains increased in these clones. The results showed that beta1,4-GalT widely affects N-gly can processing by competing with GnT-IV, GnT-V, and alpha -mannosidase II i n cells and also by some other mechanisms that suppress the conversion of h igh-mannose-type sugar chains to the hybrid type. (C) 2001 Academic Press.