Secretion of glycosylated human interleukin-2 by recombinant mammalian cell lines.
The production of glycosylated forms of the human T cell growth factor (interleukin-2, IL-2) has been studied after transfection of a mouse L cell line and a chinese hamster ovary cell line with a plasmid containing the human chromosomal interleukin-2 gene. Both cell lines produced IL-2 constitutively. Based on their behavior in reversed-phase l.c. and their sodium dodecyl sulfate-gel-electrophoresis pattern, human IL-2 protein secreted by L cells showed a similar distribution of glycosylated (Mr 16 500) and nonglycosylated (Mr 14 500) forms as the natural protein secreted by human peripheral lymphocytes, whereas the hamster cell line secreted preponderantly the glycosylated forms. Exoglycosidase digestion of the 16 500 Mr IL-2 protein shifted the gel electrophoretic mobility towards the low-molecular weight form as is true for the natural glycosylated IL-2, which contains the usual tetrasaccharide alpha-NeuAc-(2----3)-beta-D-Galp-(1----3)-[alpha-NeuAc-(2----6)]-D-GalNAc (IL-2 N2) and the trisaccharide alpha-NeuAc-(2----3)-beta-D-Galp-(1----3)-D-GalNAc (IL-2 N1) as the major carbohydrate constituents. These results support the applicability of recombinant DNA technology as a tool for studying glycoprotein biosynthesis in mammalian cells.
D000818:Animals; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004274:DNA, Recombinant / Q000378:metabolism; D005260:Female; D005796:Genes; D006026:Glycoside Hydrolases; D007376:Interleukin-2 / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D007700:Kinetics; D007739:L Cells / Q000276:immunology; D051379:Mice; D009844:Oligosaccharides / Q000032:analysis; D010053:Ovary; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism
D002241:Carbohydrates; D004274:DNA, Recombinant; D007376:Interleukin-2; D009844:Oligosaccharides; D011994:Recombinant Proteins; C050111:interleukin 2, glycosylated; D006026:Glycoside Hydrolases