Targeted disruption of Gb3/CD77 synthase gene resulted in the complete deletion of globo-series glycosphingolipids and loss of sensitivity to verotoxins

doi:10.1074/jbc.M600057200

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Papers In Press, published online ahead of print February 13, 2006
J. Biol. Chem, 10.1074/jbc.M600057200
Submitted on January 4, 2006
Revised on February 9, 2006
Accepted on February 13, 2006

Targeted disruption of Gb3/CD77 synthase gene resulted in the complete deletion of globo-series glycosphingolipids and loss of sensitivity to verotoxins

Tetsuya Okuda, Noriyo Tokuda, Shin-ichiro Numata, Masafumi Ito, Michio Ohta, Kumiko Kawamura, Joelle Wiels, Takeshi Urano, Orie Tajima, Keiko Furukawa, and Koichi Furukawa

Department of Biochemistry II, Nagoya University School of Medicine, Nagoya 466-0065

Corresponding Author: koichi{at}med.nagoya-u.ac.jp

To examine whether Gb3/CD77 is a receptor for verotoxins (VTs) in vivo, sensitivity of Gb3/CD77 synthase null mutant mice to VT-2 and VT-1 was analyzed. Although wild type mice died after the injection of 0.02 $mu$ g of VT-2 or 1.0 $mu$ g of VT-1, the mutant mice showed no reaction to even 100 times dose of individual VTs. Expression analysis of Gb3/CD77 in mouse tissues with antibody revealed that low levels but definite Gb3/CD77 was expressed in micro-vascular endothelial cells in brain cortex and piamater, and in renal tubular capillaries. Corresponding to the Gb3/CD77 expression, tissue damage with edema, congestion and cytopathic changes was observed, indicating that Gb3/CD77 (and its derivatives) exclusively function as a receptor for VTs in vivo. The lethal kinetics was almost same regardless of the elimination of LPS in VT preparation, suggesting that basal Gb3/CD77 levels are sufficient for the lethal effects of VTs.

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