Human chondroitin 6-sulfotransferase: cloning, gene structure, and chromosomal localization.
Chondroitin 6-sulfotransferase (C6ST) is the key enzyme in the biosynthesis of chondroitin 6-sulfate, a glycosaminoglycan implicated in chondrogenesis, neoplasia, atherosclerosis, and other processes. C6ST catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to carbon 6 of the N-acetylgalactosamine residues of chondroitin. Based on the previously published avian sequence, we searched the database of expressed sequence tags (dbEST) and obtained partial-length cDNAs that we completed by 5'-RACE using human chondrosarcoma and endothelial-cell RNA as template. Stable transfection of our full-length expression construct into CHO-K1 cells resulted in marked increases in C6ST and keratan sulfate sulfotransferase (KSST) enzymatic activities in cell homogenates. The predicted 411 amino acid sequence of human C6ST contains an N-terminal hydrophobic domain consistent with membrane insertion, four potential sites for N-linked glycosylation, several consensus sequences for protein phosphorylation, and one RGD sequence. The human and chick C6ST cDNA share 51% nucleotide identity, 40% amino acyl identity, and 75% amino acyl conservation. The human C6ST gene structure has been elucidated and exhibits an intron-less coding region, and the gene has been mapped to human chromosome 11 by radiation hybrid panel mapping.
Mazany
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KD
The Dorrance H. Hamilton Research Laboratories, Division of Endocrinology, Diabetes and Metabolic Diseases, Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Room 349, 1020 Locust Street, Philadelphia, PA 19107-6799, USA.
Peng
T
T
Watson
C E
CE
Tabas
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I
Williams
K J
KJ
D000116:Acetylgalactosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002807:Chondroitin / Q000378:metabolism; D002874:Chromosome Mapping; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015238:Sulfotransferases / Q000235:genetics*; D000097811:Carbohydrate Sulfotransferases
D000116:Acetylgalactosamine; D002807:Chondroitin; D018076:DNA, Complementary; D010724:Phosphoadenosine Phosphosulfate; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases