Expression cloning of a cDNA encoding a sulfotransferase involved in the biosynthesis of the HNK-1 carbohydrate epitope.
The HNK-1 carbohydrate epitope is expressed on several neural adhesion glycoproteins and as a glycolipid, and is involved in cell interactions. The structural element of the epitope common to glycoproteins and glycolipids has been determined to be sulfate-3-GlcAbeta1--> 3Galbeta1-->4GlcNAc. The glucuronyltransferase and sulfotransferase are considered to be the key enzymes in the biosynthesis of this epitope because the rest of the structure occurs often in glycoconjugates. Here we describe the isolation of the rat sulfotransferase cDNA via an expression cloning strategy. The clone finally isolated predicts a protein of 356 amino acids, with characteristics of a type II transmembrane protein and with no sequence similarity to other known sulfotransferases. Both the enzyme expressed as a soluble fusion protein and homogenates of cells transfected with the full-length cDNA could transfer sulfate from a sulfate donor to acceptor substrates containing terminal glucuronic acid.
Bakker
H
H
Department of Neurobiology, Swiss Federal Institute of Technology, Hönggerberg, 8093 Zürich, Switzerland.
Friedmann
I
I
Oka
S
S
Kawasaki
T
T
Nifant'ev
N
N
Schachner
M
M
Mantei
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018959:CD57 Antigens / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D005965:Glucuronates / Q000378:metabolism; D020723:Glucuronic Acid; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D015238:Sulfotransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D000097811:Carbohydrate Sulfotransferases
D018959:CD57 Antigens; D018076:DNA, Complementary; D005965:Glucuronates; D020723:Glucuronic Acid; D014453:Glucuronosyltransferase; D015238:Sulfotransferases; C499545:Chst10 protein, mouse; D000097811:Carbohydrate Sulfotransferases; C499547:Chst10 protein, rat; C109318:HNK-1 sulfotransferase