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dc.contributor.authorHe, Xu
dc.contributor.authorPierce, Owen
dc.contributor.authorHaselhorst, Thomas
dc.contributor.authorvon Itzstein, Mark
dc.contributor.authorKolarich, Daniel
dc.contributor.authorPacker, Nicolle H
dc.contributor.authorGloster, Tracey M
dc.contributor.authorVocadlo, David J
dc.contributor.authorQian, Yi
dc.contributor.authorBrooks, Doug
dc.contributor.authorKermode, Allison R
dc.date.accessioned2017-05-29T12:34:07Z
dc.date.available2017-05-29T12:34:07Z
dc.date.issued2013
dc.date.modified2014-03-25T22:15:53Z
dc.identifier.issn1467-7644
dc.identifier.doi10.1111/pbi.12096
dc.identifier.urihttp://hdl.handle.net/10072/57203
dc.description.abstractMucopolysaccharidosis (MPS) I is a lysosomal storage disease caused by a deficiency of a-L-iduronidase (IDUA) (EC 3.2.1.76); enzyme replacement therapy is the conventional treatment for this genetic disease. Arabidopsis cgl mutants are characterized by a deficiency of the activity of N-acetylglucosaminyl transferase I (EC 2.4.1.101), the first enzyme in the pathway of hybrid and complex N-glycan biosynthesis. To develop a seed-based platform for the production of recombinant IDUA for potential treatment of MPS I, cgl mutant seeds were generated to express human IDUA at high yields and to avoid maturation of the N-linked glycans on the recombinant human enzyme. Enzyme kinetic data showed that cgl-IDUA has similar enzymatic properties to the commercial recombinant IDUA derived from cultured Chinese hamster ovary (CHO) cells (AldurazymeTM). The N-glycan profile showed that cgl-derived IDUA contained predominantly high-mannose-type N-glycans (94.5%), and the residual complex/hybrid N-glycan-containing enzyme was efficiently removed by an additional affinity chromatography step. Furthermore, purified cgl-IDUA was amenable to sequential in vitro processing by soluble recombinant forms of the two enzymes that mediate the addition of the mannose-6-phosphate (M6P) tag in mammalian cells-UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine (GlcNAc)-1-phosphotransferase-and GlcNAc-1-phosphodiester a-N-acetylglucosaminidase (the 'uncovering enzyme'). Arabidopsis seeds provide an alternative system for producing recombinant lysosomal enzymes for enzyme replacement therapy; the purified enzymes can be subjected to downstream processing to create the M6P, a recognition marker essential for efficient receptor-mediated uptake into lysosomes of human cells.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.publisherWiley-Blackwell Publishing
dc.publisher.placeUnited Kingdom
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom1034
dc.relation.ispartofpageto1043
dc.relation.ispartofissue9
dc.relation.ispartofjournalPlant Biotechnology Journal
dc.relation.ispartofvolume11
dc.rights.retentionY
dc.subject.fieldofresearchStructural Biology (incl. Macromolecular Modelling)
dc.subject.fieldofresearchEnzymes
dc.subject.fieldofresearchTechnology
dc.subject.fieldofresearchBiological Sciences
dc.subject.fieldofresearchMedical and Health Sciences
dc.subject.fieldofresearchcode060112
dc.subject.fieldofresearchcode060107
dc.subject.fieldofresearchcode10
dc.subject.fieldofresearchcode06
dc.subject.fieldofresearchcode11
dc.titleCharacterization and downstream mannose phosphorylation of human recombinant α-L-iduronidase produced in Arabidopsis complex glycan-deficient (cgl) seeds
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2013
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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