Characterization of Sialyltransferase‐IV Activity and Its Involvement in the c‐Pathway of Brain Ganglioside Metabolism

Bettina Freischütz, Megumi Saito, Hinrich Rahmann, Robert K Yu

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Abstract: To characterize the sialyltransferase‐IV activity in brain tissues, the activities of GM1b‐, GD1a‐, GT1b‐, and GQ1c‐synthases in adult cichlid fish and rat brains were examined using GA1, GM1, GD1b, or a cod brain ganglioside mixture as the substrate. The GD1a‐synthase activity in the total membrane fraction from cichlid fish brain required divalent cations such as Mg2+ or Mn2+ and Triton CF‐54 for its full activity. The Vmax value was 1,340 pmol/mg of protein/h at an optimal pH of 6.5, whereas the apparent Km values for CMP‐sialic acid and GM1 were 172 and 78 µM, respectively. Cichlid fish and rat brains also contained GM1b‐, GT1b‐, and GQ1c‐synthase activities. The ratio of GM1b‐, GD1a‐, and GT1b‐synthase activities in fish brain was 1.00:0.89:1.13, respectively, and in rat brain 1.00:0.60:0.63. Incubation of fish brain membranes with a cod brain ganglioside mixture, which contains GT1c, and [3H]CMP‐sialic acid produced radiolabeled GQ1c. It is interesting that the adult rat brain also contains an appreciable level of GQ1c‐synthase activity despite its very low concentrations of c‐series gangliosides. The GD1a‐ or GQ1c‐synthase activity in fish and rat brain was inhibited specifically by coincubation with the glycolipids that serve as the substrates for other sialyltransferase‐IV reactions. Thus, the GD1a‐synthase activity was inhibited by GA1 and GD1b, but not by LacCer, GM3, or GD3. In a similar manner, the synthesis of GQ1c was suppressed by GA1, GM1, and GD1b, but not by LacCer, GM3, or GD3. The GD1a‐synthase activity directed toward endogenous GM1 was inhibited by GA1 or GT1b, whereas the endogenous GT1b‐synthase activity was suppressed by GA1 or GM1. GA1, GM1, and GD1b did not affect the endogenous GM3‐ and GD3‐synthase activities. These results clearly demonstrate that sialyltransferase‐IV in brain tissues catalyzes the reaction for GQ1c synthesis in the c‐pathway as well as the corresponding steps in the asialo‐, a‐, and b‐pathway in ganglioside biosynthesis.

Original languageEnglish (US)
Pages (from-to)385-393
Number of pages9
JournalJournal of Neurochemistry
Volume64
Issue number1
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Gangliosides
Metabolism
Brain
Fish
Rats
Cichlids
Gadiformes
Fishes
Tissue
Membranes
Acids
Glycolipids
Biosynthesis
Divalent Cations
Substrates

Keywords

  • Brain
  • c‐Series gangliosides
  • Fish
  • Gangliosides
  • GQ1c
  • Sialyltransferase

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Characterization of Sialyltransferase‐IV Activity and Its Involvement in the c‐Pathway of Brain Ganglioside Metabolism. / Freischütz, Bettina; Saito, Megumi; Rahmann, Hinrich; Yu, Robert K.

In: Journal of Neurochemistry, Vol. 64, No. 1, 01.01.1995, p. 385-393.

Research output: Contribution to journalArticle

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AB - Abstract: To characterize the sialyltransferase‐IV activity in brain tissues, the activities of GM1b‐, GD1a‐, GT1b‐, and GQ1c‐synthases in adult cichlid fish and rat brains were examined using GA1, GM1, GD1b, or a cod brain ganglioside mixture as the substrate. The GD1a‐synthase activity in the total membrane fraction from cichlid fish brain required divalent cations such as Mg2+ or Mn2+ and Triton CF‐54 for its full activity. The Vmax value was 1,340 pmol/mg of protein/h at an optimal pH of 6.5, whereas the apparent Km values for CMP‐sialic acid and GM1 were 172 and 78 µM, respectively. Cichlid fish and rat brains also contained GM1b‐, GT1b‐, and GQ1c‐synthase activities. The ratio of GM1b‐, GD1a‐, and GT1b‐synthase activities in fish brain was 1.00:0.89:1.13, respectively, and in rat brain 1.00:0.60:0.63. Incubation of fish brain membranes with a cod brain ganglioside mixture, which contains GT1c, and [3H]CMP‐sialic acid produced radiolabeled GQ1c. It is interesting that the adult rat brain also contains an appreciable level of GQ1c‐synthase activity despite its very low concentrations of c‐series gangliosides. The GD1a‐ or GQ1c‐synthase activity in fish and rat brain was inhibited specifically by coincubation with the glycolipids that serve as the substrates for other sialyltransferase‐IV reactions. Thus, the GD1a‐synthase activity was inhibited by GA1 and GD1b, but not by LacCer, GM3, or GD3. In a similar manner, the synthesis of GQ1c was suppressed by GA1, GM1, and GD1b, but not by LacCer, GM3, or GD3. The GD1a‐synthase activity directed toward endogenous GM1 was inhibited by GA1 or GT1b, whereas the endogenous GT1b‐synthase activity was suppressed by GA1 or GM1. GA1, GM1, and GD1b did not affect the endogenous GM3‐ and GD3‐synthase activities. These results clearly demonstrate that sialyltransferase‐IV in brain tissues catalyzes the reaction for GQ1c synthesis in the c‐pathway as well as the corresponding steps in the asialo‐, a‐, and b‐pathway in ganglioside biosynthesis.

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