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Expression cloning of a cDNA for human ceramide glucosyltransferase that catalyzes the first glycosylation step of glycosphingolipid synthesis.

May 14, 1996
93 (10) 4638-4643

Abstract

We have isolated a cDNA encoding human ceramide glucosyltransferase (glucosylceramide synthase, UDP-glucose:N-acylsphingosine D-glucosyltransferase, EC 2.4.1.80) by expression cloning using as a recipient GM-95 cells lacking the enzyme. The enzyme catalyzes the first glycosylation step of glycosphingolipid synthesis and the product, glucosylceramide, serves as the core of more than 300 glycosphingolipids. The cDNA has a G+C-rich 5' untranslated region of 290 nucleotides and the open reading frame encodes 394 amino acids (44.9 kDa). A hydrophobic segment was found near the N terminus that is the potential signal-anchor sequence. In addition, considerable hydrophobicity was detected in the regions close to the C terminus, which may interact with the membrane. A catalytically active enzyme was produced from Escherichia coli transfected with the cDNA. Northern blot analysis revealed a single transcript of 3.5 kb, and the mRNA was widely expressed in organs. The amino acid sequence of ceramide glucosyltransferase shows no significant homology to ceramide galactosyltransferase, which indicates different evolutionary origins of these enzymes.

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Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 93 | No. 10
May 14, 1996
PubMed: 8643456

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    Published online: May 14, 1996
    Published in issue: May 14, 1996

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    S Ichikawa
    Laboratory for Glyco-Cell Biology, Frontier Research Program, The Institute of Chemical and Physical Research, Saitama, Japan.
    H Sakiyama
    Laboratory for Glyco-Cell Biology, Frontier Research Program, The Institute of Chemical and Physical Research, Saitama, Japan.
    G Suzuki
    Laboratory for Glyco-Cell Biology, Frontier Research Program, The Institute of Chemical and Physical Research, Saitama, Japan.
    K I Hidari
    Laboratory for Glyco-Cell Biology, Frontier Research Program, The Institute of Chemical and Physical Research, Saitama, Japan.
    Y Hirabayashi
    Laboratory for Glyco-Cell Biology, Frontier Research Program, The Institute of Chemical and Physical Research, Saitama, Japan.

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      Expression cloning of a cDNA for human ceramide glucosyltransferase that catalyzes the first glycosylation step of glycosphingolipid synthesis.
      Proceedings of the National Academy of Sciences
      • Vol. 93
      • No. 10

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