PUBLICATION
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, T., Rouwendal, G.J., Henquet, M.G., Florack, D.E., Helsper, J.P., Bosch, D.
- ID
- ZDB-PUB-140802-3
- Date
- 2014
- Source
- Transgenic Research 23(5): 717-28 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Animals
- Biopharmaceutics/methods*
- Chickens
- Cloning, Molecular
- Galactosyltransferases/genetics*
- Galactosyltransferases/metabolism*
- Genetic Engineering/methods*
- Glycosylation
- Humans
- Nicotiana/metabolism*
- Plant Leaves/metabolism*
- Plants, Genetically Modified
- Polysaccharides/metabolism*
- Rats
- Reverse Transcriptase Polymerase Chain Reaction
- Sialyltransferases/genetics
- Species Specificity
- Zebrafish
- PubMed
- 25082356 Full text @ Transgenic. Res.
Citation
Hesselink, T., Rouwendal, G.J., Henquet, M.G., Florack, D.E., Helsper, J.P., Bosch, D. (2014) Expression of natural human β1,4-GalT1 variants and of non-mammalian homologues in plants leads to differences in galactosylation of N-glycans. Transgenic Research. 23(5):717-28.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping