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dc.contributor.authorHe, Xu
dc.contributor.authorHaselhorst, Thomas
dc.contributor.authorvon Itzstein, Mark
dc.contributor.authorKolarich, Daniel
dc.contributor.authorPacker, Nicolle H
dc.contributor.authorKermode, Allison R
dc.date.accessioned2017-05-29T12:34:03Z
dc.date.available2017-05-29T12:34:03Z
dc.date.issued2012
dc.date.modified2013-06-17T04:07:41Z
dc.identifier.issn0167-4412
dc.identifier.doi10.1007/s11103-012-9902-5
dc.identifier.urihttp://hdl.handle.net/10072/48318
dc.description.abstractProcesses associated with late events of N-glycosylation within the plant Golgi complex are a major limitation to the use of plant-based systems to produce recombinant pharmaceutical proteins for parenteral administration. Specifically, sugars added to the N-glycans of a recombinant protein during glycan maturation to complex forms (e.g. beta 1,2 xylose and alpha 1,3 fucose) can render the product immunogenic. In order to avoid these sugars, the human enzyme alpha-l-iduronidase (IDUA, EC 3.2.1.76), with a C-terminal ER-retention sequence SEKDEL, was expressed in seeds of complex-glycan-deficient (cgl) mutant and wild-type (Col-0) Arabidopsis thaliana, under the control of regulatory (5'-, signal-peptide-encoding-, and 3'-) sequences from the arcelin 5-I gene of Phaseolus vulgaris (cgl-IDUA-SEKDEL and Col-IDUA-SEKDEL, respectively). The SEKDEL motif had no adverse effect on the specific activity of the purified enzyme. Surprisingly, the majority of the N-glycans of Col-IDUA-SEKDEL were complex N-glycans (i.e. contained xylose and/or fucose) (88 %), whereas complex N-glycans comprised a much lower proportion of the N-glycans of cgl-IDUA-SEKDEL (26 %), in which high-mannose forms were predominant. In contrast to the non-chimeric IDUA of cgl seeds, which is mainly secreted into the extracellular spaces, the addition of the SEKDEL sequence to human recombinant IDUA expressed in the same background led to retention of the protein in ER-derived vesicles/compartments and its partial localization in protein storage vacuoles. Our data support the contention that the use of a C-terminal ER retention motif as an effective strategy to prevent or reduce complex N-glycan formation, is protein specific.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherSpringer
dc.publisher.placeNetherlands
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom157
dc.relation.ispartofpageto169
dc.relation.ispartofissue1-2
dc.relation.ispartofjournalPlant Molecular Biology
dc.relation.ispartofvolume79
dc.rights.retentionY
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchProtein trafficking
dc.subject.fieldofresearchGenetics
dc.subject.fieldofresearchPlant biology
dc.subject.fieldofresearchcode3101
dc.subject.fieldofresearchcode310108
dc.subject.fieldofresearchcode3105
dc.subject.fieldofresearchcode3108
dc.titleInfluence of an ER-retention signal on the N-glycosylation of recombinant human α-L-iduronidase generated in seeds of Arabidopsis
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2012
gro.hasfulltextNo Full Text
gro.griffith.authorvon Itzstein, Mark
gro.griffith.authorHaselhorst, Thomas E.
gro.griffith.authorKolarich, Daniel
gro.griffith.authorPacker, Nicki


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