Cloning, expression, and chromosomal mapping of a human ganglioside sialidase.
Here we report the cDNA sequence of a human ganglioside sialidase. The cDNA was isolated from a human brain cDNA library by screening with a 240 bp probe generated by polymerase chain reaction using primers based on the sequences of rat cytosolic and bovine membrane sialidases which we previously cloned. The 3.0 kb cDNA encodes an open reading frame of 436 amino acids containing a putative transmenbrane domain and an Arg-Ile-Pro and three Asp-box sequences characteristic of sialidases and showing overall 83% and 39% identities to the bovine and rat enzymes, respectively. Northern blot analysis revealed high expression in skeletal muscle and testis, but low level in kidney, placenta, lung, and digestive organs. Transient expression of the cDNA in COS-1 cells resulted in a 130-fold increase in sialidase activity compared to the control level, and the activity was found to be almost specific for gangliosides. Fluorescent in situ hybridization allowed the human sialidase gene localized to chromosome 11 at q 13.5.
Research Institute, Miyagi Prefectural Cancer Center, Natori, Miyagi, 981-1293, Japan.
T
T
Wada
Y
Y
Yoshikawa
S
S
Tokuyama
M
M
Kuwabara
H
H
Akita
T
T
Miyagi
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D019556:COS Cells; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000235:genetics* / Q000378:metabolism; D020161:Physical Chromosome Mapping; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014162:Transfection
C036052:GM3 ganglioside sialidase; D009439:Neuraminidase