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 language | English |
|---|---|
| Pages (from-to) | 15152-15156 |
| Number of pages | 5 |
| Journal | Journal of Biological Chemistry |
| Volume | 275 |
| Issue number | 20 |
| DOIs | |
| Publication status | Published - May 19 2000 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Molecular cloning of globotriaosylceramide/CD77 synthase, a glycosyltransferase that initiates the synthesis of globo series glycosphingolipids. / Kojima, Yoshinao; Fukumoto, Satoshi; Furukawa, Keiko; Okajima, Tetsuya; Wiels, Joelle; Yokoyama, Keiko; Suzuk, Yasuo; Urano, Takeshi; Ohta, Michio; Furukawa, Koichi.
In: Journal of Biological Chemistry, Vol. 275, No. 20, 19.05.2000, p. 15152-15156.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Molecular cloning of globotriaosylceramide/CD77 synthase, a glycosyltransferase that initiates the synthesis of globo series glycosphingolipids
AU - Kojima, Yoshinao
AU - Fukumoto, Satoshi
AU - Furukawa, Keiko
AU - Okajima, Tetsuya
AU - Wiels, Joelle
AU - Yokoyama, Keiko
AU - Suzuk, Yasuo
AU - Urano, Takeshi
AU - Ohta, Michio
AU - Furukawa, Koichi
PY - 2000/5/19
Y1 - 2000/5/19
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0034685913&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034685913&partnerID=8YFLogxK
U2 - 10.1074/jbc.M909620199
DO - 10.1074/jbc.M909620199
M3 - Article
C2 - 10748143
AN - SCOPUS:0034685913
VL - 275
SP - 15152
EP - 15156
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 20
ER -