Human N-acetylglucosamine-6-O-sulfotransferase involved in the biosynthesis of 6-sulfo sialyl Lewis X: molecular cloning, chromosomal mapping, and expression in various organs and tumor cells.

Abstract

N-Acetylglucosamine-6-O-sulfotransferase catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to position 6 of a non-reducing N-acetylglucosamine (GlcNAc) residue. We have cloned human GlcNAc-6-O-sulfotransferase cDNA, based on the sequence homology to cloned cDNA of mouse GlcNAc-6-O-sulfotransferase. The predicted protein sequence of the human enzyme was highly homologous to that of the mouse enzyme; in the 363 amino acid stretch of the catalytic region, the two proteins were nearly identical except for conservative changes in 3 amino acid residues. The expressed enzyme transferred sulfate to GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Gl cNAc. Co-transfection of the enzyme cDNA and fucosyltransferase VII cDNA into COS-7 cells resulted in cell surface expression of 6-sulfo sialyl Lewis X. Fluorescence in situ hybridization analysis revealed that the GlcNAc-6-O-sulfotransferase gene is located on human chromosome 7q31. mRNA of the human enzyme was strongly expressed in the bone marrow, peripheral blood leukocytes, spleen, brain, spinal cord, ovary, and placenta, and moderate levels of expression were observed in many organs including lymph nodes and thymus. In situ hybridization with the mouse system showed that the transcript was localized in specific regions of the brain, i.e. pyramidal cells in the CA3 subregion of the hippocampus, cerebellar nucleus and Purkinje cells. Among human tumor cells, strong expression of the mRNA was found in MOLT-4 and Jarkat lymphoblastic leukemia cells, Raji lymphoma cells, K-562 chronic myelogeneous leukemia cells, U251 glioma cells, and G361 melanoma cells. Carbohydrate structures synthesized by the sulfotransferase may be involved in various aspects of the differentiation and behavior of blood cells, their progenitor cells, and neurons in the central nervous system.

• Authors: K Uchimura, H Muramatsu, T Kaname, H Ogawa, T Yamakawa, Q W Fan, C Mitsuoka, R Kannagi, O Habuchi, I Yokoyama, K Yamamura, T Ozaki, A Nakagawara, K Kadomatsu, T Muramatsu
• Journal: Journal of biochemistry (J Biochem) Vol. 124 Issue 3 Pg. 670-8 (Sep 1998) ISSN: 0021-924X [Print] England
• PMID: 9722682 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 6'-sulfated sialyl Lewis x
• DNA, Complementary
• Lewis X Antigen
• Oligosaccharides
• RNA, Messenger
• Sialyl Lewis X Antigen
• Sulfotransferases
• carbohydrate sulfotransferases

Topics

• Amino Acid Sequence
• Animals
• Base Sequence
• Brain (metabolism)
• COS Cells
• Chromosome Mapping
• Chromosomes, Human, Pair 7
• Cloning, Molecular
• DNA, Complementary
• Humans
• In Situ Hybridization, Fluorescence
• Lewis X Antigen (analogs & derivatives)
• Mice
• Molecular Sequence Data
• Oligosaccharides (biosynthesis)
• RNA, Messenger (genetics, metabolism)
• Sequence Homology, Amino Acid
• Sialyl Lewis X Antigen (analogs & derivatives)
• Sulfotransferases (genetics, metabolism)