Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells.
The baculovirus-insect cell expression system is widely used to produce recombinant mammalian glycoproteins, but the glycosylated end products are rarely authentic. This is because insect cells are typically unable to produce glycoprotein glycans containing terminal sialic acid residues. In this study, we examined the influence of two mammalian glycosyltransferases on N-glycoprotein sialylation by the baculovirus-insect cell system. This was accomplished by using a novel baculovirus vector designed to express a mammalian alpha2,6-sialyltransferase early in infection and a new insect cell line stably transformed to constitutively express a mammalian beta1,4-galactosyltransferase. Various biochemical assays showed that a foreign glycoprotein was sialylated by this virus-host combination, but not by a control virus-host combination, which lacked the mammalian glycosyltransferase genes. Thus, this study demonstrates that the baculovirus-insect cell expression system can be metabolically engineered for N-glycoprotein sialylation by the addition of two mammalian glycosyltransferase genes.
Seo
N S
NS
Department of Molecular Biology, University of Wyoming, Laramie, Wyoming, 82071-3944, USA.
Hollister
J R
JR
Jarvis
D L
DL
D000818:Animals; D002417:Cattle; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D017781:Genes, Immediate-Early; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D009237:N-Acetyllactosamine Synthase / Q000096:biosynthesis* / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D051381:Rats; D011995:Recombination, Genetic; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000235:genetics*; D018411:Spodoptera / Q000166:cytology / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D012795:Sialoglycoproteins; D009237:N-Acetyllactosamine Synthase; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase