A genetic and structural analysis of the N-glycosylation capabilities.
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.
Léonard
Renaud
R
Department für Chemie, Universität für Bodenkultur, Muthgasse 18, A-1190 Vienna, Austria.
Kolarich
Daniel
D
Paschinger
Katharina
K
Altmann
Friedrich
F
Wilson
Iain B H
IB
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D030561:Databases, Nucleic Acid; D005647:Fucosyltransferases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010430:Pentosyltransferases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D006109:Poaceae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D014908:Triticum / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D000097608:UDP Xylose-Protein Xylosyltransferase
D005647:Fucosyltransferases; D016695:Glycosyltransferases; D010430:Pentosyltransferases; D011134:Polysaccharides; D011994:Recombinant Proteins; D000097608:UDP Xylose-Protein Xylosyltransferase