title: Characterization of a human glycoprotein (erythropoietin) produced in cultured tobacco cells.
abstract: Erythropoietin (Epo), a glycoprotein that regulates the formation of erythrocytes in mammals, was produced in cultured tobacco BY2 cells (Nicotiana tabacum L. cv. Bright Yellow 2) by introducing human Epo cDNA via Agrobacterium tumefaciens-mediated gene transfer. Epo was correctly processed and subsequently penetrated the plasma membrane of tobacco cells. However, it remained attached to the cell wall and was not released into the culture medium. Although Epo produced by tobacco cells was glycosylated with N-linked oligosaccharides, these carbohydrates were smaller than those of the recombinant Epo produced in mammalian cells. Epo produced in tobacco exhibited in vitro biological activities by inducing the differentiation and proliferation of erythroid cells. However, it had no in vivo biological activities. A lectin-binding assay indicated the lack of sialic acid residues in the N-linked oligosaccharides of Epo, suggesting that Epo was removed from the circulation before it reached erythropoietic tissues.
authors.affiliation: Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Matsumoto
authors.forename: K
authors.initials: K
authors.lastname: Ikura
authors.forename: M
authors.initials: M
authors.lastname: Ueda
authors.forename: R
authors.initials: R
authors.lastname: Sasaki
mesh_terms: D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000032:analysis; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004921:Erythropoietin / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014026:Nicotiana / Q000166:cytology / Q000235:genetics*
chemical_list: D002241:Carbohydrates; D018076:DNA, Complementary; D011994:Recombinant Proteins; D004921:Erythropoietin