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Molecular cloning and functional expression of a fifth-type alpha 2,3-sialyltransferase (mST3Gal V: GM3 synthase)

Kono, M ; Takashima, S ; Liu, H ; Inoue, M ; Kojima, N ; Lee, Y C ; Hamamoto, T ; Tsuji, S

Biochemical and biophysical research communications, 09 December 1998, Vol.253(1), pp.170-5 [Periódico revisado por pares]

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  • Título:
    Molecular cloning and functional expression of a fifth-type alpha 2,3-sialyltransferase (mST3Gal V: GM3 synthase)
  • Autor: Kono, M ; Takashima, S ; Liu, H ; Inoue, M ; Kojima, N ; Lee, Y C ; Hamamoto, T ; Tsuji, S
  • Assuntos: Sialyltransferases -- Biosynthesis
  • É parte de: Biochemical and biophysical research communications, 09 December 1998, Vol.253(1), pp.170-5
  • Descrição: The cDNA encoding a new type of alpha 2,3-sialyltransferase (mST3Gal V) was cloned from mouse brain cDNA library by PCR-based cloning approach using a pair of degenerate primers deduced from the nucleotide sequence information of mouse ST3Gal III and IV. The predicted amino acid sequence of mST3Gal V showed 27.3% and 26.4% identity to mST3Gal III and IV, respectively. The recombinant soluble mST3Gal V fused with protein-A, which expressed in the culture media of COS-7 cells, showed activity toward lactosylceramide (LacCer), and synthesized GM3. The apparent Km value for LacCer was 9.3 microM. mST3Gal V did not exhibit any activity toward other substrates we tested in this study, including glycolipids, glycoproteins and disaccharides. The mST3Gal V cDNA transfected F28-7 cells, which express large amount of lactosylceramide and very small amount of GM3 at native stage, expressed a large amount of GM3. The ST3Gal V gene was strongly expressed in mouse brain and liver, which contained a large...
  • Idioma: Inglês

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