Cited by 59Glycosylation and Glycoproteins ResearchCarbohydrate Chemistry and SynthesisCellular transport and secretion

Mouse lymphoma cell lines resistant to pea lectin are defective in fucose metabolism

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Reitman, M.L., Trowbridge, I.S., Kornfeld, S.·1980·Journal of Biological Chemistry

Abstract

Two mutants of the BW5147 mouse lymphoma cell line have been selected for their resistance to the toxic effects of pea lectin. These cell lines, termed PLR1.3 and PHAR1.8 PLR7.2, have a decreased number of high affinity pea lectin-binding sites (Trowbridge, I.S., Hyman, R., Ferson, T., and Mazauskas, C. (1978) Eur. J. Immunol. 8, 716-723). Intact cell labeling experiments using [2-3H]mannose indicated that PLR1.3 cells have a block in the conversion of GDP-[3H]mannose to GDP-[3H]fucose whereas PHAR1.8 PLR7.2 cells appear to be blocked in the transfer of fucose from GDP-[3H]fucose to glycoprotein acceptors. In vitro experiments with extracts of PLR1.3 cells confirmed the failure to convert GDP-mannose to GDP-fucose and indicated that the defect is in GDP-mannose 4,6-dehydratase (EC 4.2.1.47), the first enzyme in the conversion of GDP-mannose to GDP-fucose. The block in the PLR1.3 cells could be bypassed by growing the cells in the presence of fucose, demonstrating that an alternate pathway for the production of GDP-fucose presumably via fucose 1-phosphate is functional in this line. PLR1.3 cells grown in 10 mM fucose showed normal high affinity pea lectin binding. PHRA1.8 PLR7.2 cells synthesize GDP-fucose and have normal or increased levels of GDP-fucose:glycoprotein fucosyltransferase when assayed in vitro. The fucosyltransferases of this clone can utilize its own glycoproteins as fucose acceptors in in vitro assays. These findings indicate that this cell line fails to carry out the fucosyltransferase reaction in vivo despite the fact that it possesses the appropriate nucleotide sugar, glycoprotein acceptors, and fucosyltransferase. The finding of decreased glycoprotein fucose in two independent isolates of pea lectin-resistant cell lines and the restoration of high affinity pea lectin binding to PLR1.3 cells following fucose feeding strongly implicates fucose as a major determinant of pea lectin binding.

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Crossref · 10.1016/s0021-9258(18)43477-1

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Authors
Reitman, M. L., Trowbridge, I. S., & Kornfeld, S.
Affiliations
Salk Institute for Biological Studies (US)
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1980-10
Volume
255
Issue
20
Pages
9900-9906
ISSN
0021-9258 (ISSN-L)
OpenAlex
W1545595385
Cited by
59
References
38
Citations by year
2022: 1 · 2016: 2
FWCI
1.65
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
fucose metabolism, GDP-mannose 4,6-dehydratase, fucosyltransferase, lectin binding sites, GDP-fucose, glycoprotein synthesis
MeSH terms
Animals, Carbohydrates, Cell Line, Fucose, Fucosyltransferases, Glycopeptides, Guanosine Diphosphate Mannose, Kinetics, Lymphoma, Mutation, Lectins, Mice

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