Toward stable genetic engineering of human O-glycosylation in plants
Date
2012
Authors
Yang, Z.
Bennett, E.
Jørgensen, B.
Drew, D.
Arigi, E.
Mandel, U.
Ulvskov, P.
Levery, S.
Clausen, H.
Petersen, B.
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Journal article
Citation
Plant Physiology, 2012; 160(1):450-463
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Zhang Yang, Eric P. Bennett, Bodil Jørgensen, Damian P. Drew, Emma Arigi, Ulla Mandel, Peter Ulvskov, Steven B. Levery, Henrik Clausen, and Bent L. Petersen
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Abstract
Glycosylation is the most abundant and complex posttranslational modification to be considered for recombinant production of therapeutic proteins. Mucin-type (N-acetylgalactosamine [GalNAc]-type) O-glycosylation is found in eumetazoan cells but absent in plants and yeast, making these cell types an obvious choice for de novo engineering of this O-glycosylation pathway. We previously showed that transient implementation of O-glycosylation capacity in plants requires introduction of the synthesis of the donor substrate UDP-GalNAc and one or more polypeptide GalNAc-transferases for incorporating GalNAc residues into proteins. Here, we have stably engineered O-glycosylation capacity in two plant cell systems, soil-grown Arabidopsis (Arabidopsis thaliana) and tobacco (Nicotiana tabacum) Bright Yellow-2 suspension culture cells. Efficient GalNAc O-glycosylation of two stably coexpressed substrate O-glycoproteins was obtained, but a high degree of proline hydroxylation and hydroxyproline-linked arabinosides, on a mucin (MUC1)-derived substrate, was also observed. Addition of the prolyl 4-hydroxylase inhibitor 2,2-dipyridyl, however, effectively suppressed proline hydroxylation and arabinosylation of MUC1 in Bright Yellow-2 cells. In summary, stably engineered mammalian type O-glycosylation was established in transgenic plants, demonstrating that plants may serve as host cells for the production of recombinant O-glycoproteins. However, the present stable implementation further strengthens the notion that elimination of endogenous posttranslational modifications may be needed for the production of protein therapeutics.
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© 2012 American Society of Plant Biologists. All Rights Reserved.