Expression cloning and functional characterization of human cDNA for ganglioside GM3 synthase.
Ganglioside GM3 is a major glycosphingolipid in the plasma membrane and is widely distributed in vertebrates. We describe here the isolation of a human cDNA whose protein product is responsible for the synthesis of GM3. The cloned cDNA spanned 2,359 base pairs, with an open reading frame encoding a protein of 362 amino acids with a predicted molecular mass of 41.7 kDa. The deduced primary structure shows features characteristic of the sialyltransferase family, including a type II transmembrane topology and the sialylmotifs L at the center and S at the C-terminal region. An amino acid substitution from aspartic acid to histidine was demonstrated at a position invariant in sialylmotif L of all the other sialyltransferases so far cloned. The best acceptor substrate for the gene product was lactosylceramide, and cells transfected with the cloned cDNA clearly exhibited de novo synthesis of GM3, with a measurable decrease in the precursor lactosylceramide. Despite the ubiquitous distribution of ganglioside GM3 in human tissues, a major 2.4-kilobase transcript of the gene was found in a tissue-specific manner, with predominant expression in brain, skeletal muscle, and testis, and very low expression in liver.
Ishii
A
A
Virology and Glycobiology Division, National Cancer Center Research Institute, Chuo-ku, Tokyo 104-0045, Japan.
Ohta
M
M
Watanabe
Y
Y
Matsuda
K
K
Ishiyama
K
K
Sakoe
K
K
Nakamura
M
M
Inokuchi
J
J
Sanai
Y
Y
Saito
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D019976:Cloning, Organism; D018076:DNA, Complementary; D015723:Gene Library; D018922:HL-60 Cells; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D009928:Organ Specificity; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012799:Sialyltransferases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D011994:Recombinant Proteins; D012799:Sialyltransferases; C021343:haematoside synthetase