TAILIEUCHUNG - Báo cáo khoa học: Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants

We have compared the site-by-siteN-glycosylation status of human lactoferrin (Lf) produced in maize, a monocotyle-don, and in tobacco, usedas amodel 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 pat-tern 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 | Eur. J. Biochem. 270 3235-3242 2003 FEBS 2003 doi Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants Benedicte Samyn-Petit1. Jean-Pierre Wajda Dubos2 Frederic Chirat1. Bernadette Coddeville1 Gregory Demaizieres2 Sybille Farrer2 Marie-Christine Slomianny1 Manfred Theisen2 and Philippe Delannoy1 1 Unite de Glycobiologie Structurale et Fonctionnelle UMR CNRS 8576 Laboratoire de Chimie Biologique Universite des Sciences et Technologies de Lille Villeneuve d Ascq France 2Meristem Therapeutics Clermont-Ferrand France 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-glycosy- lation sites of each recombinant Lf are mainly substituted by typical plant paucimannose-type N-glycans with p1 2-xy-lose and a1 3-linked fucose at the proximal N-acetylgluco-samine. However tobacco Lf shows a significant amount of processed N-glycans with one or two p1 2GlcNAc linked to the trimannose core which are weakly expressed in maize Lf. Finally no Lewisa epitope was observed on tobacco Lf. Keywords glycosylation N-glycopeptides maize tobacco human lactoferrin. Several expression systems including bacteria yeast fungi insect and mammalian cells or transgenic animals are used to produce recombinant human proteins. This last decade much attention has been paid to the plant expression systems in order to express mammalian proteins. By using strong

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