Document Detail


Stable expression of human beta1,4-galactosyltransferase in plant cells modifies N-linked glycosylation patterns.
MedLine Citation:
PMID:  10200324     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
beta1,4-Galactosyltransferase (UDP galactose: beta-N-acetylglucosaminide: beta1,4-galactosyltransferase; EC 2.4.1. 22) catalyzes the transfer of galactose from UDP-Gal to N-acetylglucosamine in the penultimate stages of the terminal glycosylation of N-linked complex oligosaccharides in mammalian cells. Tobacco BY2 cells lack this Golgi enzyme. To determine to what extent the production of a mammalian glycosyltransferase can alter the glycosylation pathway of plant cells, tobacco BY2 suspension-cultured cells were stably transformed with the full-length human galactosyltransferase gene placed under the control of the cauliflower mosaic virus 35S promoter. The expression was confirmed by assaying enzymatic activity as well as by Southern and Western blotting. The transformant with the highest level of enzymatic activity has glycans with galactose residues at the terminal nonreducing ends, indicating the successful modification of the plant cell N-glycosylation pathway. Analysis of the oligosaccharide structures shows that the galactosylated N-glycans account for 47.3% of the total sugar chains. In addition, the absence of the dominant xylosidated- and fucosylated-type sugar chains confirms that the transformed cells can be used to produce glycoproteins without the highly immunogenic glycans typically found in plants. These results demonstrate the synthesis in plants of N-linked glycans with modified and defined sugar chain structures similar to mammalian glycoproteins.
Authors:
N Q Palacpac; S Yoshida; H Sakai; Y Kimura; K Fujiyama; T Yoshida; T Seki
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  96     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1999 Apr 
Date Detail:
Created Date:  1999-05-17     Completed Date:  1999-05-17     Revised Date:  2013-04-16    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  4692-7     Citation Subset:  IM    
Affiliation:
The International Center for Biotechnology, Osaka University, Yamada-oka 2-1, Suita-shi, Osaka 565, Japan.
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MeSH Terms
Descriptor/Qualifier:
Agrobacterium tumefaciens
Carbohydrate Sequence
Cell Line
Genetic Vectors
Golgi Apparatus / enzymology
Humans
Molecular Sequence Data
N-Acetyllactosamine Synthase / biosynthesis*,  genetics
Oligosaccharides / biosynthesis*,  chemistry
Plants, Toxic
Polysaccharides / biosynthesis*,  chemistry
Recombinant Proteins / biosynthesis
Tobacco
Transfection
Chemical
Reg. No./Substance:
0/Oligosaccharides; 0/Polysaccharides; 0/Recombinant Proteins; EC 2.4.1.90/N-Acetyllactosamine Synthase
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