Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation
Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. The quality and efficiency of these biopharmaceuticals are very often influenced by the glycosylation profile. Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins.
Key wordsGlycosylation Complex N-glycans Mucin-type O-glycans Glycoprotein Glycosyltransferase Glycosidases Endoplasmic reticulum Golgi apparatus Protein targeting Protein retention
We would like to thank Christiane Veit (Department of Applied Genetics and Cell Biology) for assistance in cloning and Friedrich Altmann and Daniel Maresch (both Department of Chemistry) for LC-ESI-MS-analysis. This work was supported by a grant from the Federal Ministry of Transport, Innovation and Technology (bmvit) and Austrian Science Fund (FWF): TRP 242-B20.
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