In Vivo conversion of a glycan to human compatible type by transformed tobacco cells

Kazuhito Fujiyama, Nirianne Q. Palacpac, Hiromi Sakai, Yoshinobu Kimura, Atsuhiko Shinmyo, Toshiomi Yoshida, Tatsuji Seki

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Horseradish peroxidase isozyme C (HRP; EC 1.11.1.7) was used as a model protein to evaluate the capacity of tobacco cells transformed with human β1,4-galactosyltransferase (GT6) to modify and galactosylate a foreign glycoprotein. Cells transformed with the HRP gene are designated as BY2-HRP and GT6-HRP, for wild type BY2 and GT6 transformed cells, respectively. Expression of HRP cells was confirmed by isoelectric focusing, peroxidase activity staining, Western blotting, and enzymatic assays. The presence of HRP galactosylated N-glycans in GT6-HRP cells was analyzed by lectin staining, affinity chromatography, and structural analyses of pyridylaminolabeled RCA120-bound sugar chains. The structure of Gal1GlcNAc1Man5GlcNAc2 was proposed based from the results of exoglycosidase digestions and two-dimensional sugar chain mapping. Unlike the HRP produced in BY2-HRP cells, the HRP from GT6-HRP cells has galactosylated glycoproteins that did not bind to the xylose-specific antiserum, suggesting the absence of the β1,2-xylose residue in the sugar chain.

Original languageEnglish
Pages (from-to)553-557
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume289
Issue number2
DOIs
Publication statusPublished - Nov 30 2001

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Tobacco
Sugars
Polysaccharides
Xylose
Glycoproteins
Galactosyltransferases
Affinity chromatography
Glycoside Hydrolases
Horseradish Peroxidase
Lectins
Peroxidase
Isoenzymes
Immune Sera
Assays
Genes
Staining and Labeling
Enzyme Assays
Isoelectric Focusing
Affinity Chromatography
Proteins

Keywords

  • Galactosyltransferase
  • Glycosylation
  • Tobacco

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

In Vivo conversion of a glycan to human compatible type by transformed tobacco cells. / Fujiyama, Kazuhito; Palacpac, Nirianne Q.; Sakai, Hiromi; Kimura, Yoshinobu; Shinmyo, Atsuhiko; Yoshida, Toshiomi; Seki, Tatsuji.

In: Biochemical and Biophysical Research Communications, Vol. 289, No. 2, 30.11.2001, p. 553-557.

Research output: Contribution to journalArticle

Fujiyama, Kazuhito ; Palacpac, Nirianne Q. ; Sakai, Hiromi ; Kimura, Yoshinobu ; Shinmyo, Atsuhiko ; Yoshida, Toshiomi ; Seki, Tatsuji. / In Vivo conversion of a glycan to human compatible type by transformed tobacco cells. In: Biochemical and Biophysical Research Communications. 2001 ; Vol. 289, No. 2. pp. 553-557.
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