Remodeling of sugar chain structures of human interferon-gamma.

Abstract:

:Natural human interferon (IFN)-gamma has mainly biantennary complex-type sugar chains and scarcely has multiantennary structures. We attempted to remodel the sugar chain structures using IFN-gamma as a model glycoprotein. To obtain the branching glycoforms of IFN-gamma, we introduced the genes for GnT-IV (UDP-N-acetylglucosamine:alpha-1,3-D-mannoside beta-1, 4-N-acetylglucosaminyltransferase) and/or GnT-V (UDP-N-acetylglucosamine:alpha-1,6-D-mannoside beta-1, 6-N-acetylglucosaminyltransferase) into Chinese hamster ovary (CHO) cells producing human IFN-gamma. The parental CHO cells produced IFN-gamma with biantennary sugar chains mainly. When the GnT-IV activity was increased, triantennary sugar chains with a branch produced by GnT-IV increased up to 66.9% of the total sugar chains. When the GnT-V activity was increased, triantennary sugar chains with a corresponding branch increased up to 55.7% of the total sugar chains. When the GnT-IV and -V activities were increased at a time, tetraantennary sugar chains increased up to 56.2% of the total sugar chains. The proportion of these multiantennary sugar chains corresponded to the intracellular activities of GnT-IV and -V. What is more, lectin blot and flow cytometric analysis indicated that the multi-branch structure of the sugar chains was increased not only on IFN-gamma, one of the secretory glycoproteins, but also on almost CHO cellular proteins by introducing either or both of the GnT genes. The results suggest that the branching structure of sugar chains of glycoproteins could be controlled by cellular GnT-IV and GnT-V activities. This technology can produce glycoforms out of natural occurrence, which should enlarge the potency of glycoprotein therapeutics.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Fukuta K,Abe R,Yokomatsu T,Kono N,Asanagi M,Omae F,Minowa MT,Takeuchi M,Makino T

doi

10.1093/glycob/10.4.421

subject

Has Abstract

pub_date

2000-04-01 00:00:00

pages

421-30

issue

4

eissn

0959-6658

issn

1460-2423

pii

cwd054

journal_volume

10

pub_type

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