Molecular cloning and expression of human chondroitin N-acetylgalactosaminyltransferase: the key enzyme for chain initiation and elongation of chondroitin/dermatan sulfate on the protein linkage region tetrasaccharide shared by heparin/heparan sulfate.
Based on sequence homology with the recently cloned human chondroitin synthase, we identified a novel beta1,4-N-acetylgalactosaminyltransferase, which consisted of 532 amino acids with a type II transmembrane protein topology. The amino acid sequence displayed 27% identity to that of human chondroitin synthase. The expression of a soluble form of the protein in COS-1 cells produced an active enzyme, which transferred beta1,4-N-acetylgalactosamine (GalNAc) from UDP-[(3)H]GalNAc not only to a polymer chondroitin representing growing chondroitin chains (beta-GalNAc transferase II activity) but also to GlcUAbeta1--3Galbeta1-O-C(2)H(4)NH-benzyloxycarbonyl, a synthetic substrate for beta-GalNAc transferase I that transfers the first GalNAc to the core tetrasaccharide in the protein linkage region of chondroitin sulfate. Hence, the enzyme is involved in the biosynthetic initiation and elongation of chondroitin sulfate and is the key enzyme responsible for the selective chain assembly of chondroitin/dermatan sulfate on the linkage region tetrasaccharide common to various proteoglycans containing chondroitin/dermatan sulfate or heparin/heparan sulfate chains. The coding region of this enzyme was divided into seven discrete exons and localized to chromosome 8. Northern blot analysis revealed that the chondroitin GalNAc transferase gene exhibited a ubiquitous but markedly differential expression in human tissues and that the expression pattern was similar to that of chondroitin synthase. Thus, more than two distinct enzymes forming the novel gene family are required for chain initiation and elongation in chondroitin/dermatan sulfate as in the biosynthesis of heparin/heparan sulfate.
Uyama
Toru
T
Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
Kitagawa
Hiroshi
H
Tamura Ji
Jun-ichi
J
Sugahara
Kazuyuki
K
D000116:Acetylgalactosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D002809:Chondroitin Sulfates / Q000096:biosynthesis*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003871:Dermatan Sulfate / Q000096:biosynthesis*; D014453:Glucuronosyltransferase; D016695:Glycosyltransferases / Q000235:genetics; D006493:Heparin / Q000096:biosynthesis*; D006497:Heparan Sulfate / Q000096:biosynthesis*; D006801:Humans; D008969:Molecular Sequence Data; D064251:Multifunctional Enzymes; D017350:N-Acetylgalactosaminyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D000116:Acetylgalactosamine; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D014453:Glucuronosyltransferase; D016695:Glycosyltransferases; D006493:Heparin; D006497:Heparan Sulfate; D064251:Multifunctional Enzymes; D017350:N-Acetylgalactosaminyltransferases; C412725:chondroitin synthase; C498800:CHSY1 protein, human