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Title: Molecular Mechanism of Substrate Specificity for Heparan Sulfate 2-O-Sulfotransferase

Authors:
; ; ; ; ; ; ; ;  [1];  [2];  [2]
  1. (Shandong)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1140042
Resource Type:
Journal Article
Resource Relation:
Journal Name: J. Biol. Chem.; Journal Volume: 289; Journal Issue: (19) ; 05, 2014
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Liu, Chunhui, Sheng, Juzheng, Krahn, Juno M., Perera, Lalith, Xu, Yongmei, Hsieh, Po-Hung, Dou, Wenfang, Liu, Jian, Pedersen, Lars C., NIH), and UNC). Molecular Mechanism of Substrate Specificity for Heparan Sulfate 2-O-Sulfotransferase. United States: N. p., 2014. Web. doi:10.1074/jbc.M113.530535.
Liu, Chunhui, Sheng, Juzheng, Krahn, Juno M., Perera, Lalith, Xu, Yongmei, Hsieh, Po-Hung, Dou, Wenfang, Liu, Jian, Pedersen, Lars C., NIH), & UNC). Molecular Mechanism of Substrate Specificity for Heparan Sulfate 2-O-Sulfotransferase. United States. doi:10.1074/jbc.M113.530535.
Liu, Chunhui, Sheng, Juzheng, Krahn, Juno M., Perera, Lalith, Xu, Yongmei, Hsieh, Po-Hung, Dou, Wenfang, Liu, Jian, Pedersen, Lars C., NIH), and UNC). Fri . "Molecular Mechanism of Substrate Specificity for Heparan Sulfate 2-O-Sulfotransferase". United States. doi:10.1074/jbc.M113.530535.
@article{osti_1140042,
title = {Molecular Mechanism of Substrate Specificity for Heparan Sulfate 2-O-Sulfotransferase},
author = {Liu, Chunhui and Sheng, Juzheng and Krahn, Juno M. and Perera, Lalith and Xu, Yongmei and Hsieh, Po-Hung and Dou, Wenfang and Liu, Jian and Pedersen, Lars C. and NIH) and UNC)},
abstractNote = {},
doi = {10.1074/jbc.M113.530535},
journal = {J. Biol. Chem.},
number = (19) ; 05, 2014,
volume = 289,
place = {United States},
year = {Fri Jul 11 00:00:00 EDT 2014},
month = {Fri Jul 11 00:00:00 EDT 2014}
}
  • Heparan sulfate (HS) 3-O-sulfotransferase isoform-2 (3-OST-2), which belongs to a family of enzymes capable of generating herpes simplex virus type-1 (HSV-1) entry and spread receptors, is predominantly expressed in human brain. Despite its unique expression pattern, the ability of 3-OST-2 to mediate HSV-1 entry and cell-to-cell fusion is not known. Our results demonstrate that expression of 3-OST-2 can render Chinese hamster ovary K1 (CHO-K1) cells susceptible to entry of wild-type and mutant strains of HSV-1. Evidence for generation of gD receptors by 3-OST-2 were suggested by gD-mediated interference assay and the ability of 3-OST-2-expressing CHO-K1 cells to preferentially bind HSV-1more » gD, which could be reversed by prior treatment of cells with HS lyases (heparinases II/III). In addition, 3-OST-2-expressing CHO-K1 cells acquired the ability to fuse with cells-expressing HSV-1 glycoproteins, a phenomenon that mimics a way of viral spread in vivo. Demonstrating specificity, the cell fusion was inhibited by soluble 3-O-sulfated forms of HS, but not unmodified HS. Taken together, our results raise the possibility of a role of 3-OST-2 in the spread of HSV-1 infection in the brain.« less
  • Heparin and heparan sulfate are linear sulfated polysaccharides that exert a multitude of biological functions. Heparan sulfate glucosaminyl N-deacetylase/N-sulfotransferase isoform 2 (NDST-2), a key enzyme in the biosynthesis of heparin, contains two distinct activities. This bifunctional enzyme removes the acetyl group from N-acetylated glucosamine (N-deacetylase activity) and transfers a sulfuryl group to the unsubstituted amino position (N-sulfotransferase activity). The N-sulfotransferase activity of NDST has been unambiguously localized to the C-terminal domain of NDST. Here, we report that the N-terminal domain of NDST-2 retains N-deacetylase activity. The N-terminal domain (A66-P604) of human NDST-2, designated as N-deacetylase (NDase), was cloned as amore » (His){sub 6}-fusion protein, and protein expression was carried out in Escherichia coli. Heparosan treated with NDase contains N-unsubstituted glucosamine and is highly susceptible to N-sulfation by N-sulfotransferase. Our results conclude that the N-terminal domain of NDST-2 contains functional N-deacetylase activity. This finding helps further elucidate the mechanism of action of heparan sulfate N-deacetylase/N-sulfotransferases and the biosynthesis of heparan sulfate in general.« less
  • The phase 2 metabolic reactions, sulfation and glucuronidation, were studied in a human colon carcinoma cell line (Caco-2), which has been developed as a model of intestinal enterocytes. Phenol sulfotransferase was isolated from Caco-2 cells cultured for 7, 14 and 21 days. The enzyme catalyzed the sulfation of both p-nitrophenol and catecholamines as well as most catecholamine metabolites. The affinity (K{sub m}) of PST for dopamine was much higher than for p-nitrophenol, and the specific activity of PST with both substrates increased with the age of the cells. The thermal stability of Caco-2 PST increased with cell age and wasmore » not dependent on the acceptor substrate used. The thermolabile PST from 7-day old cells was more sensitive to NEM than was the thermostable enzyme from 21-day old cells. No UDP-glucuronyltransferase activity was detected in 7-, 14- and 21-day old Caco-2 cells with any of the methods used.« less
  • Treacher Collins syndrome is an autosomal dominant disorder of craniofacial development, the features of which include conductive hearing loss and cleft palate. Previous studies have shown that the Treacher Collins syndrome locus is flanked by D5S519 and SPARC, and a yeast artificial chromosome contig encompassing this {open_quotes}critical region{close_quotes} has been completed. In the current investigation a cosmid containing D5S519 has been used to screen a human placental cDNA library. This has resulted in the cloning of the human heparan sulfate-N-deacetylase/N-sulfotransferase gene. Two different mRNA species that have identical protein coding sequences but that differ in the size and sequence ofmore » the 3{prime} untranslated regions (3{prime}UTR) have been identified. The smaller species has a 3{prime}UTR of 1035 bp, whereas that of the larger is 4878 bp. 24 refs., 3 figs.« less