Introduction of tri-antennary N-glycans in Arabidopsis thaliana plants

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• 作者：Bieke Nagels^a ; ^b ; ^{Bieke.Nagels@UGent.be} ; Els J.M. Van Damme^a ; ^{ElsJM.VanDamme@UGent.be} ; Nico Callewaert^c ; ^d ; ^{Nico.Callewaert@UGent.be} ; Koen Weterings^b ; ¹ ; ^{Koen.Weterings@bayer.com}
• 关键词：Arabidopsis thaliana ; N-acetylglucosaminyltransferase ; N-acetylhexosaminidase ; N-glycosylation ; Tri-antennary
• 刊名：Plant Science
• 出版年：2012
• 出版时间：May 2012
• 年：2012
• 卷：185-186
• 期：Complete
• 页码：161-168
• 全文大小：697 K

文摘

Because the pathway for protein synthesis is largely conserved between plants and animals, plants provide an attractive platform for the cost effective and flexible production of biopharmaceuticals. However, there are some differences in glycosylation between plants and humans that need to be considered before plants can be used as an efficient expression platform.

In the presented research the human genes encoding ¦Á1,3-mannosyl-¦Â1,4-N-acetylglucosaminyltransferase (GnT-IV) and ¦Á1,6-mannosyl-¦Â1,6-N-acetylglucosaminyltransferase (GnT-V) were introduced in the fast cycling model plant Arabidopsis thaliana to synthesize tri-antennary N-glycans. The GnT-IV and -V enzymes were targeted to the Golgi apparatus with plant-specific localization signals. The experiments were performed both in a wild type background, as well as in plants lacking ¦Â1,2-xylosyltransferase (XylT) and ¦Á1,3-fucosyltransferase (FucT) activity. Glycan analysis of endogenous proteins in the transgenic lines using CE-LIF showed that tri-antennary N-glycans could be produced in the XylT/FucT deficient line, while these structures were not found in the wild type background. Since ¦Â-N-acetylhexosaminidases, that remove terminal GlcNAcs, are active in A. thaliana plants, the specificity of these enzymes for different GlcNAc linkages was tested. The results showed that there is no pronounced preference of the A. thaliana hexosaminidases for human-type GlcNAc-linkages.