Title : Engineering of N. benthamiana L. plants for production of N-acetylgalactosamine-glycosylated proteins--towards development of a plant-based platform for production of protein therapeutics with mucin type O-glycosylation.

Pub. Date : 2010 Aug 24

PMID : 20735851






7 Functional Relationships(s)
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1 RESULTS: The present study demonstrates that an isoform of the human GalNAc-Ts family, GalNAc-T2, retains its localization and functionality upon expression in N. benthamiana L. plants. N-acetylgalactosaminuronic acid polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
2 A GalNAc-T2-specific acceptor peptide, the 113-136 aa fragment of chorionic gonadotropin beta-subunit, is glycosylated in vitro by the plant-produced enzyme at the "native" GalNAc attachment sites, Ser-121 and Ser-127. Serine polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
3 A GalNAc-T2-specific acceptor peptide, the 113-136 aa fragment of chorionic gonadotropin beta-subunit, is glycosylated in vitro by the plant-produced enzyme at the "native" GalNAc attachment sites, Ser-121 and Ser-127. Serine polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
4 Ectopic expression of GalNAc-T2 is sufficient to "arm" tobacco cells with the ability to perform GalNAc-glycosylation, as evidenced by the attachment of GalNAc to Thr-119 of the endogenous enzyme endochitinase. N-acetylgalactosaminuronic acid polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
5 Ectopic expression of GalNAc-T2 is sufficient to "arm" tobacco cells with the ability to perform GalNAc-glycosylation, as evidenced by the attachment of GalNAc to Thr-119 of the endogenous enzyme endochitinase. Threonine polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
6 Further genetic engineering of the GalNAc-T2 plants by co-expressing Y. enterocolitica UDP-GlcNAc 4-epimerase gene and C. elegans UDP-GlcNAc/UDP-GalNAc transporter gene overcomes these limitations as indicated by the expression of the model LTBMUC1 protein exclusively as a glycoform. 2-acetamido-2-deoxy-4-O-(beta-2-acetamid-2-deoxyglucopyranosyl)glucopyranose polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens
7 Further genetic engineering of the GalNAc-T2 plants by co-expressing Y. enterocolitica UDP-GlcNAc 4-epimerase gene and C. elegans UDP-GlcNAc/UDP-GalNAc transporter gene overcomes these limitations as indicated by the expression of the model LTBMUC1 protein exclusively as a glycoform. Uridine Diphosphate N-Acetylglucosamine polypeptide N-acetylgalactosaminyltransferase 2 Homo sapiens