Molecular cloning and expression of a glycosaminoglycan N-acetylglucosaminyl N-deacetylase/N-sulfotransferase from a heparin-producing cell line
Department of Biochemistry and Molecular Biology
Amidohydrolases; Amino Acid Sequence; Animals; Base Sequence; Cell Line; Cloning, Molecular; DNA, Complementary; *Gene Expression; Heparin; Kinetics; Liver; Mast-Cell Sarcoma; Molecular Sequence Data; RNA, Messenger; Rats; Sequence Homology, Amino Acid; Sulfotransferases; Transfection; Tumor Cells, Cultured
Life Sciences | Medicine and Health Sciences
Heparin has a higher content of N-sulfated glucosamine and L-iduronic acid than heparan sulfate. Deacetylation of N-acetylglucosamine followed by N-sulfation may be important steps differentiating the biosynthesis of these glycosaminoglycans. We have cloned, by cross-hybridization with the cDNA from rat liver heparan sulfate N-deacetylase/N-sulfotransferase, a protein from a heparin synthesizing mastocytoma derived cell line called MST. This protein, which has both N-deacetylase/N-sulfotransferase activities, has a predicted amino acid sequence homology of 70% with the above rat liver enzyme and is unique for the following reasons. 1) It was found to be encoded by a 3.8-kilobase mRNA that was unique to heparin-producing cells; an 8.5-kilobase mRNA encoding the rat liver enzymes has been found to occur in all mammalian cells tested on the basis of nucleic acid cross-hybridization; 2) the protein overexpressed in COS cells in its full-length transmembrane form or as a soluble secreted protein A chimera displayed ratios of N-deacetylase to N-sulfotransferase activities that were 4-8-fold higher than that observed for the enzyme found in liver that is involved in the biosynthesis of heparan sulfate. These results suggest that the MST-derived enzyme is probably unique to the production of heparin in mast cells.
J Biol Chem. 1994 Jan 21;269(3):2270-6.
The Journal of biological chemistry
Orellana A, Hirschberg CB, Wei Z, Swiedler SJ, Ishihara M. (1994). Molecular cloning and expression of a glycosaminoglycan N-acetylglucosaminyl N-deacetylase/N-sulfotransferase from a heparin-producing cell line. Open Access Publications by UMass Chan Authors. Retrieved from https://escholarship.umassmed.edu/oapubs/827