title: Expression of natural human β1,4-GalT1 variants and of non-mammalian homologues in plants leads to differences in galactosylation of N-glycans.
abstract: β1,4-Galactosylation of plant N-glycans is a prerequisite for commercial production of certain biopharmaceuticals in plants. Two different types of galactosylated N-glycans have initially been reported in plants as the result of expression of human β1,4-galactosyltransferase 1 (GalT). Here we show that these differences are associated with differences at its N-terminus: the natural short variant of human GalT results in hybrid type N-glycans, whereas the long form generates bi-antennary complex type N-glycans. Furthermore, expression of non-mammalian, chicken and zebrafish GalT homologues with N-termini resembling the short human GalT N-terminus also induce hybrid type N-glycans. Providing both non-mammalian GalTs with a 13 amino acid N-terminal extension that distinguishes the two naturally occurring forms of human GalT, acted to increase the levels of bi-antennary galactosylated N-glycans when expressed in tobacco leaves. Replacement of the cytosolic tail and transmembrane domain of chicken and zebrafish GalTs with the corresponding region of rat α2,6-sialyltransferase yielded a gene whose expression enhanced the level of bi-antennary galactosylation even further.
authors.affiliation: Plant Research International B.V., Wageningen University and Research Center, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands, thamara.hesselink@wur.nl.
authors.forename: Thamara
authors.initials: T
authors.lastname: Hesselink
authors.forename: Gerard J A
authors.initials: GJ
authors.lastname: Rouwendal
authors.forename: Maurice G L
authors.initials: MG
authors.lastname: Henquet
authors.forename: Dion E A
authors.initials: DE
authors.lastname: Florack
authors.forename: Johannes P F G
authors.initials: JP
authors.lastname: Helsper
authors.forename: Dirk
authors.initials: D
authors.lastname: Bosch
mesh_terms: D000818:Animals; D001702:Biopharmaceutics / Q000379:methods*; D002645:Chickens; D003001:Cloning, Molecular; D005700:Galactosyltransferases / Q000235:genetics* / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D018515:Plant Leaves / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011134:Polysaccharides / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012799:Sialyltransferases / Q000235:genetics; D013045:Species Specificity; D014026:Nicotiana / Q000378:metabolism*; D015027:Zebrafish; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
chemical_list: D011134:Polysaccharides; D005700:Galactosyltransferases; C493310:beta1,4-galactosyltransferase, human; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
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