Information on EC 5.1.3.17 - heparosan-N-sulfate-glucuronate 5-epimerase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
5.1.3.17
-
RECOMMENDED NAME
GeneOntology No.
heparosan-N-sulfate-glucuronate 5-epimerase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Epimerization of D-glucuronate in heparosan-N-sulfate to L-iduronate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
epimerization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycosaminoglycan biosynthesis - heparan sulfate / heparin
-
-
heparan sulfate biosynthesis (late stages)
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-
Metabolic pathways
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-
SYSTEMATIC NAME
IUBMB Comments
poly[(1->4)-beta-D-glucuronosyl-(1->4)-N-sulfo-alpha-D-glucosaminyl] glucurono-5-epimerase
The enzyme acts on D-glucosyluronate residues in N-sulfated heparosan polymers, converting them to L-iduronate, thus modifying the polymer to heparan-N-sulfate. The enzyme requires that at least the N-acetylglucosamine residue linked to C-4 of the substrate has been deacetylated and N-sulfated, and activity is highest with fully N-sulfated substrate. It does not act on glucuronate residues that are O-sulfated or are adjacent to N-acetylglucosamine residues that are O-sulfated at the 6 position. Thus the epimerization from D-glucuronate to L-iduronate occurs after N-sulfation of glucosamine residues but before O-sulfation. Not identical with EC 5.1.3.19 chondroitin-glucuronate 5-epimerase or with EC 5.1.3.36, heparosan-glucuronate 5-epimerase.
CAS REGISTRY NUMBER
COMMENTARY hide
112567-86-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene Hsepi
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
C57BL/6 mice
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
type 15
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
GlcA-beta-(1->4)-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Octa-1, two GlcA units in Octa-1 are susceptible to C5-epi modification, and both epimerization sites are reversible
i.e. epi-Octa-1
-
r
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Octa-4
i.e. epi-Octa-4
-
ir
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Octa-3, two GlcA units in Octa-1 are susceptible to C5-epi modification, and both epimerization sites are reversible
i.e. epi-Octa-3
-
ir
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Deca-8 with GlcA-beta-(1->4)-GlcNAc-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA(irreversible site)-beta-(1->4)-GlcNS-beta-(1->4)-GlcA (reversible site)-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
i.e. epi-Deca-8
-
?
GlcA-beta-(1->4)-GlcNH2-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNH2-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Octa-6
i.e. epi-Octa-6
-
r
GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Hexa-7
i.e. epi-Hexa-7
-
r
GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
GlcA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-IdoA-beta-(1->4)-GlcNS-beta-(1->4)-GlcA-beta-(1->4)-2,5-andydro-D-mannitol
show the reaction diagram
-
i.e. Octa-2
i.e. epi-Octa-2
-
r
heparosan N-sulfate D-glucuronate
heparosan N-sulfate L-iduronate
show the reaction diagram
-
-
-
-
?
Heparosan-N-sulfate D-glucuronate
Heparosan-N-sulfate L-iduronate
show the reaction diagram
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
heparosan N-sulfate D-glucuronate
heparosan N-sulfate L-iduronate
show the reaction diagram
-
-
-
-
?
Heparosan-N-sulfate D-glucuronate
Heparosan-N-sulfate L-iduronate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
-
additional information
-
no specific metal ion requirement
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KCl
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stimulates, maximal effect at 0.1 M, inhibition at higher concentrations
NaCl
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stimulates, maximal effect at 0.05 M, inhibition at higher concentrations
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2
heparan sulfate-derived polysaccharide
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-
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0.17
heparin-derived polysaccharide
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0.063 - 0.44
uronic acid residues
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
620
uronic acid residues
Bos taurus
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-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
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7
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.1 - 7.9
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pH 6.1: about 60% of maximal activity, pH 7.9: about 30% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
; midbrain and hindbrain
Manually annotated by BRENDA team
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decreased GLCE expression is observed in 10% of benign prostate hyperplasia tissues and 53% of prostate tumors, and increased GLCE mRNA levels are detected in 49% of benign prostate hyperplasia tissues and 21% of tumors, intratumoral heterogeneity of GLCE protein levels both in benign prostate hyperplasia and prostate cancer cells, resulting in a mixed population of GLCE-expressing and nonexpressing epithelial cells in vivo
Manually annotated by BRENDA team
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decreased GLCE expression is observed in 10% of benign prostate hyperplasia tissues and 53% of prostate tumors, and increased GLCE mRNA levels are detected in 49% of benign prostate hyperplasia tissues and 21% of tumors, intratumoral heterogeneity of GLCE protein levels both in benign prostate hyperplasia and prostate cancer cells, resulting in a mixed population of GLCE-expressing and nonexpressing epithelial cells in vivo
Manually annotated by BRENDA team
additional information
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quantitative RT-PCR enzyme expression analysis in benign and malign prostate tissues, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
transmembrane enzyme
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
49905
x * 49905, calculation from nucleotide sequence
51914
x * 51914, sequence calculation, x * 52000, recombinant enzyme, SDS-PAGE
52000 - 56000
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gel filtration
54000
-
1 * 54000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer or tetramer
x * 51914, sequence calculation, x * 52000, recombinant enzyme, SDS-PAGE
monomer
-
1 * 54000, SDS-PAGE
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
purified recombinant His-tagged enzyme, inactivation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 10% glycerol, retains full activity for 1.5 years
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli
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recombinant His-tagged enzyme from Escherichia coli strain Rossetta(DE3)/pWUR537 by cobalt affinity chromatography
recombinant wild-type and mutant enzymes from Escherichia coli strain DE3
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned into a baculovirus expression vector and expressed in Sf9 insect cells
expressed in U2020 cells
-
expression of wild-type and mutant enzymes in Escherichia coli strain DE3
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gene Hsepi
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gene RED65_08024, design and synthesis of a synthetic based on hypothetical protein RED65_08024, codon optimized for the codon usage bias of Escherichia coli, expression of the recombinant His-tagged enzyme in Escherichia coli strain Rossetta(DE3)/pWUR537
recombinant expression in Escherichia coli
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recombinant full-length enzyme, expressed in insect Sf9 cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
D-glucuronyl C5-epimerase expression is decreased in lung cancer cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Y146A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y162A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y168A
-
site-directed mutagenesis, inactive mutant
Y210A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y222A
-
site-directed mutagenesis, inactive mutant
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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assay procedure for heparosan-N-sulfate-glucuronate 5-epimerase, which is based on the use of a two-phase system for liquid scintillation counting
medicine