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J Biol Chem, Vol. 273, Issue 41, 26729-26738, October 9, 1998

Expression Cloning and Characterization of a Novel Murine alpha 1,3-Fucosyltransferase, mFuc-TIX, That Synthesizes the Lewis x (CD15) Epitope in Brain and Kidney

Takashi KudoDagger , Yuzuru IkeharaDagger , Akira TogayachiDagger , Mika KanekoDagger §, Tsuneo HiragaDagger , Katsutoshi Sasaki, and Hisashi NarimatsuDagger

From the Dagger  Division of Cell Biology, Institute of Life Science, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, the § Department of Genetics, School of Life Science, The Graduate University for Advanced Studies, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-0801, and the  Tokyo Research Laboratories, Kyowa Hakko Kogyo Company, Limited, 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan

The 3-fucosyl-N-acetyllactosamine (Lewis x, CD15, SSEA-1) carbohydrate epitope is widely distributed in many tissues and is developmentally expressed in some rodent and human tissues, i.e. brain and lung, and mouse early embryo. In such tissues, the Lewis x epitope is considered to be involved in cell-cell interactions. We isolated a novel mouse alpha 1,3-fucosyltransferase gene, named mFuc-TIX, from an adult mouse brain cDNA library using the expression cloning method. On flow cytometric analysis, Namalwa cells transfected stably with the mFuc-TIX gene showed a marked increase in Lewis x epitopes but not sialyl Lewis x epitopes. As seen experiments involving oligosaccharides as acceptor substrates, mFuc-TIX transfers a fucose to lacto-N-neotetraose but not to either alpha 2,3-sialyl lacto-N-neotetraose or lacto-N-tetraose. The substrate specificity of mFuc-TIX was similar to that of mouse myeloid-type alpha 1,3-fucosyltransferase (mFuc-TIV). The deduced amino acid sequence of mFuc-TIX, consisting of 359 residues, indicated a type II membrane protein and shows low degrees of homology to the previously cloned alpha 1,3-fucosyltransferases, i.e. mFuc-TIV (48.4%), mouse Fuc-TVII (39.1%), and human Fuc-TIII (43.0%), at the amino acid sequence level. A phylogenetic tree of the alpha 1,3-fucosyltransferases constructed by the neighbor-joining method showed that mFuc-TIX is quite distant from the other alpha 1,3-fucosyltransferases. Thus, mFuc-TIX does not belong to any subfamilies of known alpha 1,3Fuc-Ts. The mFuc-TIX transcript was mainly detected in brain and kidney with the Northern blotting and competitive reverse transcription-polymerase chain reaction methods, whereas the mFuc-TIV transcript was not detected in brain with these methods. On in situ hybridization, the mFuc-TIX transcript was detected in neuronal cells but not in the glial cells including astrocytes. These results strongly indicated that mFuc-TIX participates in the Lewis x synthesis in neurons of the brain and may be developmentally regulated.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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