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Volume 270,
Number 15,
Issue of April 14, pp. 8844-8850, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Molecular
Cloning and Expression of Two Types of Rabbit -Galactoside
1,2-Fucosyltransferase
Seiji
Hitoshi
,
Susumu
Kusunoki
,
Ichiro
Kanazawa
,
Shuichi
Tsuji
Two DNA clones encoding rabbit -galactoside
1,2-fucosyltransferase (RFT-I and RFT-II) have been isolated from
a rabbit genomic DNA library. The DNA sequences revealed open reading
frames coding for 373 (RFT-I) and 354 (RFT-II) amino acids,
respectively. The deduced amino acid sequences of RFT-I and RFT-II
showed 56% identity with each other, and that of RFT-I showed 80%
identity with that of human H blood type 1,2-fucosyltransferase.
Northern blot analysis of embryo and adult rabbit tissues revealed that
the RFT-I gene was expressed in adult brain, and that the RFT-II gene
was expressed in salivary and lactating mammary glands. The identities
of these enzymes were confirmed by constructing recombinant
fucosyltransferases in which the N-terminal part including the
cytoplasmic tail and signal anchor domain was replaced with the
immunoglobulin signal peptide sequence. RFT-I expressed in COS-7 cells
exhibited similar transferase activity to that of human H blood type
1,2-fucosyltransferase. RFT-II expressed in COS-7 cells showed
higher affinity for type 1 (Gal 1,3GlcNAc) and type 3
(Gal 1,3GalNAc) acceptors than type 2 (Gal 1,4GlcNAc) ones,
which suggested that RFT-II was a putative secretor-type
1,2-fucosyltransferase.

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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.
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