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https://hdl.handle.net/2440/76418
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Type: | Journal article |
Title: | Engineering mammalian mucin-type O-glycosylation in plants |
Author: | Yang, Z. Drew, D. Jorgensen, B. Mandel, U. Bach, S. Ulvskov, P. Levery, S. Bennett, E. Clausen, H. Petersen, B. |
Citation: | Journal of Biological Chemistry, 2012; 287(15):11911-11923 |
Publisher: | Amer Soc Biochemistry Molecular Biology Inc |
Issue Date: | 2012 |
ISSN: | 0021-9258 1083-351X |
Statement of Responsibility: | Zhang Yang, Damian P. Drew, Bodil Jørgensen, Ulla Mandel, Søren S. Bach, Peter Ulvskov, Steven B. Levery, Eric P. Bennett, Henrik Clausen, and Bent L. Petersen |
Abstract: | Mucin-type O-glycosylation is an important post-translational modification that confers a variety of biological properties and functions to proteins. This post-translational modification has a particularly complex and differentially regulated biosynthesis rendering prediction and control of where O-glycans are attached to proteins, and which structures are formed, difficult. Because plants are devoid of GalNAc-type O-glycosylation, we have assessed requirements for establishing human GalNAc O-glycosylation de novo in plants with the aim of developing cell systems with custom-designed O-glycosylation capacity. Transient expression of a Pseudomonas aeruginosa Glc(NAc) C4-epimerase and a human polypeptide GalNAc-transferase in leaves of Nicotiana benthamiana resulted in GalNAc O-glycosylation of co-expressed human O-glycoprotein substrates. A chimeric YFP construct containing a 3.5 tandem repeat sequence of MUC1 was glycosylated with up to three and five GalNAc residues when co-expressed with GalNAc-T2 and a combination of GalNAc-T2 and GalNAc-T4, respectively, as determined by mass spectrometry. O-Glycosylation was furthermore demonstrated on a tandem repeat of MUC16 and interferon α2b. In plants, prolines in certain classes of proteins are hydroxylated and further substituted with plant-specific O-glycosylation; unsubstituted hydroxyprolines were identified in our MUC1 construct. In summary, this study demonstrates that mammalian type O-glycosylation can be established in plants and that plants may serve as a host cell for production of recombinant O-glycoproteins with custom-designed O-glycosylation. The observed hydroxyproline modifications, however, call for additional future engineering efforts. |
Keywords: | Glycoprotein Biosynthesis Mucins Plant Protein Expression Vaccine Development GalNAc Glycoengineering Nicotiana benthamiana O-Glycosylation |
Rights: | © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. |
DOI: | 10.1074/jbc.M111.312918 |
Published version: | http://dx.doi.org/10.1074/jbc.m111.312918 |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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