Cloned (Comment) | Organism |
---|---|
gene NDST1, recombinant enzyme expression | Homo sapiens |
gene NDST4, recombinant expression of C-terminally His6-tagged full-length NDST-4 and of luminal domain of NDST4 (Ser35-Arg872) in Spodoptera frugiperda Sf9 cells using the insect baculovirus transfection system | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
Golgi apparatus | - |
Homo sapiens | 5794 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine | Homo sapiens | - |
adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P52848 | - |
- |
Homo sapiens | Q9H3R1 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | four potential N-glycosylation sites are present in NDST-4, making the glycosylation become one potential reason why recombinant NDST-4 lacks N-deacetylase activity. The baculovirus expression system is capable of transferring N-linked oligosaccharide side chains to the same sites in recombinant proteins as in the native protein in mammalian cells | Homo sapiens |
Purification (Comment) | Organism |
---|---|
recombinant enzyme by heparin affinity chromatography | Homo sapiens |
recombinant His6-tagged NDST-4 by nickel affinity and heparin affinity chromatography | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3'-phosphoadenylyl sulfate + GlcA-GlcNAc-GlcA-GlcNS-GlcA-pNP | - |
Homo sapiens | adenosine 3',5'-bisphosphate + ? | - |
? | |
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine | - |
Homo sapiens | adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine | - |
? | |
additional information | substrate specificity of recombinant NDST-1 (rNDST-1), full-length enzyme and individual N-deacetylase and N-sulfotransferase domains, with model oligosaccharides, detailed overview. Saccharides N-sulfated by NDST-1 are highly ordered, alternately containing N-S domains and N-Ac domains. Domain cooperation plays an essential role in NDST-1 production of HS with specific structures, and N-deacetylation is the limiting step. Recombinant NDST-1 clearly displayed bifunctional activity, it shows N-deacetylase and N-sulfotransferase activity, and the minimum-sized substrate for the bifunctional activity is a hexasaccharide | Homo sapiens | ? | - |
- |
|
additional information | substrate specificity of recombinant NDST-4 (rNDST-4) with model oligosaccharides, detailed overview. Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. NDST-4 does not show directional N-sulfotransferase activity and the N-deacetylase domain is inactive. rNDST-4 modified heparosan only contained below 5% disaccharide units with the structure of GlcA-GlcNS. rNDST-4 displays low N-deacetylase activity toward polysaccharide. rNDST-4 is not active with a heptasaccharide substrate or smaller | Homo sapiens | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
? | x * 110000, recombinant rNDST-4 with signal peptide, SDS-PAGE | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
heparan sulfate N-deacetylase/N-sulfotransferase isoform 1 | - |
Homo sapiens |
heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 | - |
Homo sapiens |
NDST | - |
Homo sapiens |
NDST-1 | - |
Homo sapiens |
NDST-4 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | four NDST isoforms are present in humans | Homo sapiens |
evolution | four NDST isoforms are present in the human. NDST-1 is the most widely distributed isoform in tissues and organisms, and is considered to exhibit profound physiological functions during all stages of human development | Homo sapiens |
additional information | analysis of the mode of action of NDST-4 | Homo sapiens |
additional information | NDST-1 has two catalytic domains | Homo sapiens |
physiological function | NDSTs are potential bifunctional enzymes that contain two independent catalytic domains: the N-deacetylase domain removes the acetyl group from GlcNAc to form GlcNH2, and the N-sulfo-transferase domain transfers a sulfo group to the GlcNH2 residue to form GlcNS. The enzyme is involved in the synthesis of heparan sulfate (HS), a highly sulfated linear glycosaminoglycan polysaccharide, consisting of a disaccharide repeating unit of sulfated derivatives of glucosamine and glucuronic acid (GlcA) or iduronic acid (IdoA) residues. HS is widely present in proteoglycan saccharide chains in mammalian tissues and the extracellular matrix. The final structure of HS chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. N-Deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotranferases. NDST-4 plays a limited role in heparan sulfate biosynthesis | Homo sapiens |
physiological function | NDSTs are potential bifunctional enzymes that contain two independent catalytic domains: the N-deacetylase domain removes the acetyl group from GlcNAc to form GlcNH2, and the N-sulfo-transferase domain transfers a sulfo group to the GlcNH2 residue to form GlcNS. The enzyme is involved in the synthesis of heparan sulfate (HS), a highly sulfated linear glycosaminoglycan polysaccharide, consisting of a disaccharide repeating unit of sulfated derivatives of glucosamine and glucuronic acid (GlcA) or iduronic acid (IdoA) residues. HS is widely present in proteoglycan saccharide chains in mammalian tissues and the extracellular matrix. The final structure of HS chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. N-Deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotranferases | Homo sapiens |