Accelerated biosynthesis of neolacto-series glycosphingolipids in differentiated mouse embryonal carcinoma F9 cells detected by using dodecyl N-acetylglucosaminide as a saccharide primer

Nao Ogasawara, Yohko U. Katagiri, Nobutaka Kiyokawa, Tomonori Kaneko, Ban Sato, Hideki Nakajima, Yoshitaka Miyagawa, Yasunori Kushi, Hideharu Ishida, Makoto Kiso, Hajime Okita, Toshinori Sato, Junichiro Fujimoto

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Using dodecyl N-acetylglucosaminide (GlcNAc-C12) as a saccharide primer, we investigated the biosynthetic changes of neolacto-series glycosphingolipids (GSLs) in mouse embryonal carcinoma F9 cells during differentiation induced by retinoic acid plus dibutyryl cyclic AMP (RA/dbcAMP). In the differentiated cells, the glycosylation of GlcNAc-C12 was greatly enhanced. The sugar compositions of glycosylated primers were assigned as Hex-GlcNAc, [Hex]2-GlcNAc, [Hex]2[HexNAc]-GlcNAc, and [NeuAc][Hex]-GlcNAc by liquid chromatography-tandem mass spectrometry. The detection of augmented biosynthesis of endogenous sialylparagloboside indicated that [NeuAc][Hex]-GlcNAc was predicted to be the non-reducing end trisaccharide of sialylparagloboside. The transcription of B3gnt5, B4galt1, Ggta1, Fut4 and St3gal6, encoding glycosyltransferases involved in the neolacto-series glycosphingolipids biosynthesis, was increased, whereas that of Fut9 and St6galI was decreased after RA/dbcAMP treatment. Furthermore, the sialyltransferase activity of ST3GalVI sialylating paragloboside was enhanced with the increase in St3gal6 expression. Since most stage-specific embryonic antigen-1 (SSEA-1) active determinants are carried by glycoproteins in F9 cells, the changes in glycolipid metabolism do not seem to be closely related to loss of cell surface SSEA-1 expression upon F9 differentiation. These results indicate that RA/dbcAMP treatment activates the biosynthesis of neolacto-series GSL and enhances sialylation of paragloboside in F9 cells with down-regulation of Fut9 expression.

Original languageEnglish
Pages (from-to)321-330
Number of pages10
JournalJournal of biochemistry
Volume149
Issue number3
DOIs
Publication statusPublished - 2011 Mar 1

Keywords

  • F9
  • Fut9
  • GlcNAc-C12 primer
  • St3gal6
  • sialylparagloboside

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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