Document Detail


N-glycosylation of a baculovirus-expressed recombinant glycoprotein in three insect cell lines.
MedLine Citation:
PMID:  9542646     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The capacity of two Trichoplusia ni (TN-368 and BTI-Tn-5b 1-4) and a Spodoptera frugiperda (IPLB-SF-21A) cell lines to glycosylate recombinant, baculovirus-encoded, secreted, placental alkaline phosphatase was compared. The alkaline phosphatase from serum-containing, cell culture medium was purified by phosphate affinity column chromatography. The N-linked oligosaccharides were released from the purified protein with PNGase F and analyzed by fluorophore-assisted carbohydrate electrophoresis. The majority of oligosaccharide structures produced by the three cell lines contained two or three mannose residues, with and without core fucosylation, but there were structures containing up to seven mannose residues. The oligosaccharides that were qualitatively or quantitatively different between the cell lines were sequenced with glycosidase digestions. The S. frugiperda cells produced more fucosylated oligosaccharides than either of the T. ni cell lines. The smallest oligosaccharide produced by S. frugiperda cells was branched trimannose. In contrast, both T. ni cell lines produced predominantly dimannose and linear trimannose structures devoid of alpha 1-3-linked mannose.
Authors:
P C Kulakosky; M L Shuler; H A Wood
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  In vitro cellular & developmental biology. Animal     Volume:  34     ISSN:  1071-2690     ISO Abbreviation:  In Vitro Cell. Dev. Biol. Anim.     Publication Date:  1998 Feb 
Date Detail:
Created Date:  1998-05-21     Completed Date:  1998-05-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9418515     Medline TA:  In Vitro Cell Dev Biol Anim     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  101-8     Citation Subset:  IM    
Affiliation:
Boyce Thompson Institute for Plant Research, Ithaca, New York 14853, USA.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / analysis*,  genetics
Animals
Carbohydrate Sequence
Cell Line
Genetic Vectors*
Glycoproteins / analysis*,  genetics
Glycosylation
Molecular Sequence Data
Moths
Nucleopolyhedrovirus*
Oligosaccharides / analysis*
Placenta / enzymology*
Recombinant Fusion Proteins / analysis,  genetics
Spodoptera
Chemical
Reg. No./Substance:
0/Glycoproteins; 0/Oligosaccharides; 0/Recombinant Fusion Proteins; EC 3.1.3.1/Alkaline Phosphatase

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