Molecular cloning of globotriaosylceramide/CD77 synthase, a glycosyltransferase that initiates the synthesis of globo series glycosphingolipids

Yoshinao Kojima, Satoshi Fukumoto, Keiko Furukawa, Tetsuya Okajima, Joelle Wiels, Keiko Yokoyama, Yasuo Suzuk, Takeshi Urano, Michio Ohta, Koichi Furukawa

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

The expression cloning of a cDNA for globotriaosylceramide (Gb3)/CD77 synthase (α1,4-galactosyltransferase) was achieved using an anti-Gb3 antibody and mouse L cells as a recipient cell line for the transfection. The isolated cDNA clone designated pVTR1 predicted a type II membrane protein with 19 amino acids of cytoplasmic domain, 26 amino acids of transmembrane region, and a catalytic domain with 308 amino acids. Introduction of the cDNA clone into L cells resulted in the neosynthesis of Gb3/CD77, and the extracts of the transfectant cells showed α1,4-galactosyltransferase activity only on lactosylceramide and galactosylceramide. In Northern blotting, a 2.3-kilobase mRNA was strongly expressed in heart, kidney, spleen, and placenta and weakly in colon, small intestine, and brain. Transfection of the cDNA into L cells resulted in the constitution of sensitivity to the apoptosis with Shiga-like toxins (verotoxins). Since Gb3/CD77 synthase initiates the synthesis of globo series glycolipids, the isolation of this cDNA will make possible further investigations into the function of its important series of glycolipids.

Original languageEnglish
Pages (from-to)15152-15156
Number of pages5
JournalJournal of Biological Chemistry
Volume275
Issue number20
DOIs
Publication statusPublished - May 19 2000
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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