Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants.
We have compared the site-by-site N-glycosylation status of human lactoferrin (Lf) produced in maize, a monocotyledon, and in tobacco, used as a model dicotyledon. Maize and tobacco plants were stably transformed and recombinant Lf was purified from both seeds and leaves. N-glycopeptides were generated by trypsin digestion of recombinant Lf and purified by reverse-phase HPLC. The N-glycosylation pattern of each site was determined by mass spectrometry. Our results indicated that the N-glycosylation patterns of recombinant Lf produced in maize and tobacco share common structural features. In particular, both N-glycosylation sites of each recombinant Lf are mainly substituted by typical plant paucimannose-type N-glycans, with beta1,2-xylose and alpha1,3-linked fucose at the proximal N-acetylglucosamine. However, tobacco Lf shows a significant amount of processed N-glycans with one or two beta1,2GlcNAc linked to the trimannose core, which are weakly expressed in maize Lf. Finally, no Lewisa epitope was observed on tobacco Lf.
Samyn-Petit
Bénédicte
B
Unité de Glycobiologie Structurale et Fonctionnelle, UMR CNRS 8576, Laboratoire de Chimie Biologique, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Wajda Dubos
Jean-Pierre
JP
Chirat
Frédéric
F
Coddeville
Bernadette
B
Demaizieres
Grégory
G
Farrer
Sybille
S
Slomianny
Marie-Christine
MC
Theisen
Manfred
M
Delannoy
Philippe
P
D001216:Asparagine / Q000032:analysis; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D007781:Lactoferrin / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic; D014357:Trypsin / Q000378:metabolism; D003313:Zea mays / Q000235:genetics*
D006020:Glycopeptides; D011994:Recombinant Proteins; D001216:Asparagine; D007781:Lactoferrin; D014357:Trypsin