RRC ID 39101
Author Fukuta M, Inazawa J, Torii T, Tsuzuki K, Shimada E, Habuchi O.
Title Molecular cloning and characterization of human keratan sulfate Gal-6-sulfotransferase.
Journal J Biol Chem
Abstract We have previously cloned chondroitin 6-sulfotransferase (C6ST) cDNA from chick embryo chondrocytes. C6ST catalyzes sulfation of chondroitin, keratan sulfate, and sialyl N-acetyllactosamine oligosaccharides. In this study, we report the cloning and characterization of a novel sulfotransferase that catalyzes sulfation of keratan sulfate. This new sulfotransferase cDNA clone was obtained from a human fetal brain library by cross-hybridization with chick C6ST cDNA. The cDNA clone obtained contains a single open reading frame that predicts a type II transmembrane protein composed of 411 amino acid residues. When the cDNA was introduced into a eukaryotic expression vector and transfected in COS-7 cells, keratan sulfate sulfotransferase activity was overexpressed, but C6ST activity was not increased over that of the control. Structural analysis of 35S-labeled glycosaminoglycan, which was formed from keratan sulfate by the reaction with 35S-labeled 3'-phosphoadenosine 5'-phosphosulfate and the recombinant sulfotransferase, showed that keratan sulfate was sulfated at position 6 of Gal residues. On the basis of the acceptor substrate specificity, we propose keratan sulfate Gal-6-sulfotransferase (KSGal6ST) for the name of the newly cloned sulfotransferase. KSGal6ST was assigned to chromosome 11p11. 1-11.2 by fluorescence in situ hybridization. Among various human adult tissues, a 2.8-kilobase message of KSGal6ST was expressed mainly in the brain. When poly(A)+ RNAs from the chick embryo cornea and brain were probed with the human KSGal6ST cDNA in Northern hybridization, a clear band with about 2.8 kilobases was detected. These observations suggest that KSGal6ST may participate in the biosynthesis of keratan sulfate in the brain and cornea.
Volume 272(51)
Pages 32321-8
Published 1997-12-19
DOI 10.1074/jbc.272.51.32321
PII S0021-9258(18)39767-9
PMID 9405439
MeSH Amino Acid Sequence Animals Base Sequence Blotting, Northern COS Cells Chromatography, High Pressure Liquid Chromosome Mapping Chromosomes, Human, Pair 11 Cloning, Molecular DNA, Complementary Humans In Situ Hybridization, Fluorescence Molecular Sequence Data Sequence Homology, Amino Acid Substrate Specificity Sulfotransferases / genetics* Sulfotransferases / isolation & purification Sulfotransferases / metabolism
IF 4.238
Times Cited 113
Human and Animal Cells COS-7(RCB0539)