Document Detail


Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells.
MedLine Citation:
PMID:  11437599     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
N S Seo; J R Hollister; D L Jarvis
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Protein expression and purification     Volume:  22     ISSN:  1046-5928     ISO Abbreviation:  Protein Expr. Purif.     Publication Date:  2001 Jul 
Date Detail:
Created Date:  2001-07-04     Completed Date:  2001-12-07     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  United States    
Other Details:
Languages:  eng     Pagination:  234-41     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
Cell Line, Transformed
Cloning, Molecular
Genes, Immediate-Early
Genetic Vectors / chemistry,  genetics,  isolation & purification
N-Acetyllactosamine Synthase / biosynthesis*,  genetics*
Nucleopolyhedrovirus / genetics*
Rats
Recombination, Genetic
Sialoglycoproteins / biosynthesis*,  genetics
Sialyltransferases / biosynthesis*,  genetics*
Spodoptera / cytology,  enzymology,  genetics*,  metabolism*
Grant Support
ID/Acronym/Agency:
GM49734/GM/NIGMS NIH HHS; R01 GM049734/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Sialoglycoproteins; EC 2.4.1.90/N-Acetyllactosamine Synthase; EC 2.4.99.-/Sialyltransferases; EC 2.4.99.1/beta-D-galactoside alpha 2-6-sialyltransferase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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