Involvement of murine β-1,4-galactosyltransferase v in lactosylceramide biosynthesis

Tadahiro Kumagai, Takeshi Sato, Shunji Natsuka, Yukito Kobayashi, Dapeng Zhou, Tadashi Shinkai, Satoru Hayakawa, Kiyoshi Furukawa

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32 Citations (Scopus)

Abstract

Human β-1,4-galactosyltransferase (β-1,4-GalT) V was shown to be involved in the biosynthesis of N-glycans, O-glycans and lactosylceramide (Lac-Cer) by in vitro studies. To determine its substrate specificity, enzymatic activity and its products were analyzed using mouse embryonic fibroblast (MEF) cells from β-1,4-GalT V (B4galt5)-mutant mice. Analysis of expression levels of the β-1,4-GalT I-VI genes revealed that the expression of the β-1,4-GalT V gene in B4galt5 +/- - and B4galt5 -/- -derived MEF cells are a half and null when compared to that of B4galt5 +/+ -derived MEF cells without altering the expression levels of other β-1,4-GalT genes. These MEF cells showed no apparent difference in their growth. When β-1,4-GalT activities were determined towards GlcNAcβ-S-pNP, no significant difference in its specific activity was obtained among B4galt5 +/+ -, B4galt5 +/- - and B4galt5 -/- -derived MEF cells. No significant differences were obtained in structures and amounts of N-glycans and lectin bindings to membrane glycoproteins among B4galt5 +/+ -, B4galt5 +/- - and B4galt5 -/- -derived MEF cells. However, when cell homogenates were incubated with glucosylceramide in the presence of UDP-[3H]Gal, Lac-Cer synthase activity in B4galt5 +/- - and B4galt5 -/- -derived MEF cells decreased to 41% and 11% of that of B4galt5 +/+ -derived MEF cells. Consistent with this, amounts of Lac-Cer and its derivative GM3 in B4galt5 -/- -derived MEF cells decreased remarkably when compared with those of B4galt5 +/+ -derived MEF cells. These results indicate that murine β-1,4-GalT V is involved in Lac-Cer biosynthesis.

Original languageEnglish
Pages (from-to)685-695
Number of pages11
JournalGlycoconjugate Journal
Volume27
Issue number7-9
DOIs
Publication statusPublished - 2010 Oct 1

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Keywords

  • B4galt5 mice
  • Lactosylceramide
  • MEF cells
  • β-1,4-GalT V

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kumagai, T., Sato, T., Natsuka, S., Kobayashi, Y., Zhou, D., Shinkai, T., Hayakawa, S., & Furukawa, K. (2010). Involvement of murine β-1,4-galactosyltransferase v in lactosylceramide biosynthesis. Glycoconjugate Journal, 27(7-9), 685-695. https://doi.org/10.1007/s10719-010-9313-2