Information on EC 2.8.2.8 - [heparan sulfate]-glucosamine N-sulfotransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.8.2.8
-
RECOMMENDED NAME
GeneOntology No.
[heparan sulfate]-glucosamine N-sulfotransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine = adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
sulfate group transfer
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
heparan sulfate biosynthesis (late stages)
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SYSTEMATIC NAME
IUBMB Comments
3'-phosphoadenylyl-sulfate:[heparan sulfate]-glucosamine N-sulfotransferase
The enzyme also catalyses the sulfation of chondroitin 4-sulfate and dermatan sulfate, but to a much more limited extent.
CAS REGISTRY NUMBER
COMMENTARY hide
9026-75-9
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
possesses only 1 isozyme
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-
Manually annotated by BRENDA team
enzyme form NDST1 and NDST2, but not NDST3; wild-type and mutant pgsE-606
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Manually annotated by BRENDA team
possesses only 1 isozyme
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Manually annotated by BRENDA team
hen
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
isozymes NDST1-4
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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the absence of both isoforms Ndst1 and Ndst2 induces increased branching of the ductal epithelium in mammary gland. Ndst-deficient ducts display an apparent overabundance of branched ductal epithelia and cellular chimerism in the level of sulfated cell surface heparan sulfate
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3'-phosphoadenylyl sulfate + (heparan sulfate)-glucosamine
adenosine 3',5'-bisphosphate + (heparan sulfate)-N-sulfoglucosamine
show the reaction diagram
NDST-1 plays a role in lung development and repair following injury due to its influence on the activity of fibroblast growth factor
-
-
?
3'-phosphoadenylyl sulfate + heparosan
adenosine 3',5'-bisphosphate + heparosan sulfate
show the reaction diagram
3'-phosphoadenylyl sulfate + N-desulfated heparin
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
-
-
-
-
?
3'-phosphoadenylyl sulfate + [heparan sulfate]-D-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfo-D-glucosamine
show the reaction diagram
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-D-sulfoglucosamine
show the reaction diagram
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine
show the reaction diagram
3'-phosphoadenylylsulfate + chondroitin 4-sulfate
?
show the reaction diagram
-
weak activity
-
-
?
3'-phosphoadenylylsulfate + de-N-sulfated heparin
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
-
-
-
-
?
3'-phosphoadenylylsulfate + dermatan sulfate
?
show the reaction diagram
3'-phosphoadenylylsulfate + desulfated N-acetylated heparan sulfate
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
-
-
-
-
?
3'-phosphoadenylylsulfate + heparan sulfate
adenosine 3',5'-bisphosphate + N-sulfoheparan sulfate
show the reaction diagram
3'-phosphoadenylylsulfate + heparin
adenosine 3',5'-bisphosphate + N-sulfoheparin
show the reaction diagram
3'-phosphoadenylylsulfate + N,O-desulfated heparan sulfate
adenosine 3',5'-bisphosphate + O-desulfated heparan sulfate
show the reaction diagram
3'-phosphoadenylylsulfate + N,O-desulfoheparan sulfate tetrasaccharides with the nonreducing terminus occupied by glucuronic acid
?
show the reaction diagram
3'-phosphoadenylylsulfate + N-acetyl heparosan
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
-
-
-
-
?
3'-phosphoadenylylsulfate + N-acetylated heparan sulfate
?
show the reaction diagram
3'-phosphoadenylylsulfate + N-deacetylated K5 polysaccharide
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
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-
-
-
?
3'-phosphoadenylylsulfate + N-deacetylated K5-polysaccharide
?
show the reaction diagram
3'-phosphoadenylylsulfate + N-desulfated heparan sulfate
adenosine 3',5'-bisphosphate + heparan sulfate
show the reaction diagram
3'-phosphoadenylylsulfate + N-desulfated heparin
?
show the reaction diagram
3'-phosphoadenylylsulfate + N-desulfated, N-acetylated heparin
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
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poor substrate
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-
?
3'-phosphoadenylylsulfate + N-desulfoheparin
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
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-
-
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?
3'-phosphoadenylylsulfate + oligosaccharides derived from N-desulfoheparan sulfate
?
show the reaction diagram
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-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
3'-phosphoadenylyl sulfate + (heparan sulfate)-glucosamine
adenosine 3',5'-bisphosphate + (heparan sulfate)-N-sulfoglucosamine
show the reaction diagram
Q02353
NDST-1 plays a role in lung development and repair following injury due to its influence on the activity of fibroblast growth factor
-
-
?
3'-phosphoadenylyl sulfate + [heparan sulfate]-D-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfo-D-glucosamine
show the reaction diagram
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-D-sulfoglucosamine
show the reaction diagram
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine
adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine
show the reaction diagram
3'-phosphoadenylylsulfate + N-desulfoheparin
adenosine 3',5'-bisphosphate + ?
show the reaction diagram
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?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3',5'-ADP
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CNMQALSMPVTC
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the peptide inhibits isoform Ndst1 activity by direct interaction with the enzyme near the active site, the peptide inhibits sulfotransferase activity by 25 and 40% at 0.01 and 0.1 mM, respectively
CRGWRGEKIGNC
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the peptide blocks the binding of substrate to the enzyme isoform Ndst1
Cu2+
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NMQALSMPVT
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PCMB
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phenylmercuric acetate
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RGWRGEKIGN
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Zn2+
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-
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
estrogen
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enhances activity with N,O-desulfated heparan sulfate as acceptor, progesterone suppresses the effect of estrogen
Heparin sulfate
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excess heparin sulfate increases the N-sulfotransferase activity of heparin sulfate-modifying N-deacetylase/N-sulfotransferase
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005 - 0.108
3'-phosphoadenylylsulfate
2.5
N,O-desulfoheparan sulfate tetrasaccharide
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calculated from disaccharide units
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0.0009 - 0.0224
N-deacetylase K5-polysaccharide
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1.89
N-desulfoheparan sulfate
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calculated from disaccharide units
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0.013
N-desulfoheparin
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apparent value, in 50 mM HEPES, pH 7.0, at 37C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.14
NMQALSMPVT
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apparent value, in 50 mM HEPES, pH 7.0, at 37C
0.48
RGWRGEKIGN
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apparent value, in 50 mM HEPES, pH 7.0, at 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.95
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substrate heparan sulfate
231
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purified recombinant sulfotransferase domain, substrate is at all positions completely desulfated heparin
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2
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6.3
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assay at
6.7 - 7.2
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.7 - 8.1
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pH 5.7: 33% of activity maximum, pH 8.1: 54% of activity maximum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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clone E6-1, ATCC TIB-152
Manually annotated by BRENDA team
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gene Ndst1 is expressed during lens development, in lens epithelium, strong expression occurs in both the lens placode and in the optic vesicle in mouse embryos at stage E9.5, overview
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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high expression level of isozyme NDST2
Manually annotated by BRENDA team
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high expression level of isozyme NDST2. The expression of NDST2 in monocytes is modified following cell activation or maturation, although the same treatment has no significant effect on the level of mRNAencoding NDST1
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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CHO cells
Manually annotated by BRENDA team
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CD4+ cells, high expression level of isozyme NDST2. NDST1 is barely detected in naive T-lymphocytes by comparison with memory and activated CD4 T cells, whereas no notable variation in the expression of NDST2 can be distinguished between T cell subsets
Manually annotated by BRENDA team
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promonocytic leukemia cells
Manually annotated by BRENDA team
embryonic, cell culture
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
92000
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radiation inactivation analysis
94000
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1 * 94000, SDS-PAGE
97000
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gel filtration
additional information
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peaks with enzyme activity: 200000 and 110000 Da
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 94000, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
18 mg/ml, 0.1 M Tris-HCl, pH 8.3, 0.1 M NaCl, 4 mM 3'-phosphoadenosine 5'-phosphate, reservoir solution: Bis-Tris-propane, pH 7.0, 0.2 M NaCl, 30% PEG 3000, room temperature, X-ray diffraction structure determination and analysis; purified recombinant bacterially expressed sulfotransferase domain
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18 mg/ml, 0.1 M Tris-HCl, pH 8.3, 0.1 M NaCl, 4 mM 3'-phosphoadenosine 5'-phosphate, reservoir solution: Bis-Tris-propane, pH 7.0, 0.2 M NaCl, 30% PEG 3000, room temperature, X-ray diffraction structure determination and analysis; purified recombinant selenomethionyl sulfotransferase domain NST1
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structure
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
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2 min, no effect
50
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1 min, 15% loss of activity
85
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2 min, complete loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18C, 20 h, stable
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-18C, stable for at least 4 months
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-70C, stable for at least 6 months in presence of 20% glycerol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
heparin-Toyopearl column chromatography and nickel-agarose column chromatography
IgG-Sepharose bead chromatography
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N-acetylglucosaminyl N-deacetylase/N-sulfotransferase, recombinant from COS-1 cells, 1-step affinity chromatographical purification
N-desulfo-N-acetylheparan sulfate deacetylase activity copurifies
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recombinant fusion tagged sulfotransferase domain from Escherichia coli BL21 cells by affinity chromatography
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recombinant sulfotransferase domain NST1 from Escherichia coli strain B834 (DE3)
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
brain cDNA library, DNA sequence determination and analysis; construction of a chimeric enzyme consisiting of a soluble enzyme form NDST3 N-terminally fused to a protein A fragment, expression in COS-7 cells
DNA and RNA determination in hepatocytes
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DNA sequence determination and analysis, genetic structure and organization
DNA sequence determination, analysis, and expression of the wild-type and mutated sulfotransferase domain of the bifunctional enzyme in Escherichia coli BL21 cells as fusion tagged protein
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expressed in HEK-293 cells
expressed in pgsA745 cells
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expressed in Saccharomyces cerevisiae InvSc1 cells
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expressed in Sf9 insect cells
expression in Saccharomyces cerevisiae
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expression of the bifunctional enzyme heparan sulfate/heparin N-deacetylase/N-sulfotransferase-1 in Saccharomyces cerevisiae, yeast-expressed enzyme has both N-deacetylase and N-sulfotransferase activities
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expression of the selenomethionyl sulfotransferase domain NST1 in methionine auxotrophic Escherichia coli strain B834 (DE3)
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isozyme NDST1, DNA sequence determination and analysis; isozyme NDST2, DNA sequence determination and analysis; isozyme NDST3, DNA sequence determination and analysis; isozyme NDST4, DNA sequence determination and analysis
isozyme NDST3, chromosomal localisation; isozyme NDST4, DNA sequence determination and analysis, chromosomal localization
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isozymes NDST1 and NDST2
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lambdagt10 library construction and screening, DNA sequence determination and analysis, expression in COS-1 cells
overexpression of isozymes NDST1 and NDST2 in human kidney cells 293; stable functional overexpression in human kidney cell line 293, 2.5fold increase in N-sulfation and 8fold increase of N-deacetylation of glucosaminyl residues of human heparan sulfate in transfected cells
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overexpression of the bifunctional enzyme in CHO cells
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stable functional overexpression in human kidney cell line 293, 2.5fold increase in N-sulfation and 8fold increase of N-deacetylation of glucosaminyl residues of human heparan sulfate in transfected cells
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the N-terminal domain (A66-P604) of the bifunctional enzyme heparan sulfate N-deacetylase/N-sulfotransferase 2 is cloned as a (His)6-fusion protein, expression in Escherichia coli. The N-terminal domain contains functional N-deacetylase activity
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
NDST-2 is downregulated after mast cell activation, e.g. by calcium ionophore or IgE ligation. Down-regulation of NDST-2 gene by A23187
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K614A
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site-directed mutagenesis of the sulfotransferase domain of the bifunctional enzyme, complete loss of N-sulfotransferase activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
medicine
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diabetes substantially suppresses hepatic N-deacetylase/N-sulfotransferase-1 mRNA, protein and enzymatic activity. Suppression of hepatic N-deacetylase/N-sulfotransferase-1 may contribute to diabetic dyslipidemia. Stimulation of N-deacetylase/N-sulfotransferase-1 activity by AngII inhibitors may provide cardiovascular protection