Abstract
The recent draft sequencing of the rice (Oryza sativa) genome has enabled a genetic analysis of the glycosylation capabilities of an agroeconomically important group of plants, the monocotyledons. In this study, we have not only identified genes putatively encoding enzymes involved in N-glycosylation, but have examined by MALDI-TOF MS the structures of the N-glycans of rice and other monocotyledons (maize, wheat and dates; Zea mays, Triticum aestivum and Phoenix dactylifera); these data show that within the plant kingdom the types of N-glycans found are very similar between monocotyledons, dicotyledons and gymnosperms. Subsequently, we constructed expression vectors for the key enzymes forming plant-typical structures in rice, N-acetylglucosaminyltransferase I (GlcNAc-TI; EC 2.4.1.101), core alpha1,3-fucosyltransferase (FucTA; EC 2.4.1.214) and beta1,2-xylosyltransferase (EC 2.4.2.38) and successfully expressed them in Pichia pastoris. Rice GlcNAc-TI, FucTA and xylosyltransferase are therefore the first monocotyledon glycosyltransferases involved in N-glycan biosynthesis to be characterised in a recombinant form.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Carbohydrate Sequence
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Databases, Nucleic Acid
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Fucosyltransferases / analysis
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Fucosyltransferases / genetics
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Fucosyltransferases / metabolism
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Glycosylation
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Glycosyltransferases / analysis
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Glycosyltransferases / genetics
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Glycosyltransferases / metabolism
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Molecular Sequence Data
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Oryza / enzymology*
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Oryza / genetics
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Oryza / metabolism
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Pentosyltransferases / analysis
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Pentosyltransferases / genetics
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Pentosyltransferases / metabolism
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Pichia / genetics
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Poaceae / enzymology
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Poaceae / genetics
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Poaceae / metabolism
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Polysaccharides / analysis
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Polysaccharides / biosynthesis*
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Recombinant Proteins / analysis
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Substrate Specificity
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Triticum / enzymology
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Triticum / genetics
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Triticum / metabolism
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UDP Xylose-Protein Xylosyltransferase
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Zea mays / enzymology
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Zea mays / genetics
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Zea mays / metabolism
Substances
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Polysaccharides
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Recombinant Proteins
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Glycosyltransferases
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Fucosyltransferases
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Pentosyltransferases
Associated data
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GENBANK/AJ457976
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GENBANK/AJ582955
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GENBANK/AJ621917
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GENBANK/AJ621918