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- Title
Expression of natural human β1,4-GalT1 variants and of non-mammalian homologues in plants leads to differences in galactosylation of N-glycans.
- Authors
Hesselink, Thamara; Rouwendal, Gerard; Henquet, Maurice; Florack, Dion; Helsper, Johannes; Bosch, Dirk
- 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.
- Subjects
GLYCOSYLATION; GLYCANS; GENE expression in plants; BIOPHARMACEUTICS; GALACTOSYLTRANSFERASE genetics
- Publication
Transgenic Research, 2014, Vol 23, Issue 5, p717
- ISSN
0962-8819
- Publication type
Article
- DOI
10.1007/s11248-014-9806-z