5.1.3.17: heparosan-N-sulfate-glucuronate 5-epimerase
This is an abbreviated version!
For detailed information about heparosan-N-sulfate-glucuronate 5-epimerase, go to the full flat file.
Word Map on EC 5.1.3.17
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5.1.3.17
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l-iduronic
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d-glucuronic
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epimerization
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proteoglycans
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idoa
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glycosaminoglycans
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glca
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n-sulfated
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2-o-sulfotransferase
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hexuronic
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hspgs
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2-o-sulfation
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mastocytoma
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mannuronan
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lindahl
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o-desulfated
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medicine
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diagnostics
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analysis
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synthesis
- 5.1.3.17
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l-iduronic
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d-glucuronic
-
epimerization
- proteoglycans
-
idoa
- glycosaminoglycans
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glca
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n-sulfated
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2-o-sulfotransferase
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hexuronic
-
hspgs
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2-o-sulfation
- mastocytoma
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mannuronan
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lindahl
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o-desulfated
- medicine
- diagnostics
- analysis
- synthesis
Reaction
Synonyms
C5 uronosyl epimerase, C5-epi, C5-epimerase, D-Glucuronyl C-5 epimerase, D-glucuronyl C5-epimerase, Epimerase, polyglucuronate, Glce, GlceA, GlceB, glucuronosyl C-5 epimerase, glucuronyl C5-epimerase, heparan sulfate C-5 epimerase, Heparan sulfate C5-epimerase, Heparosan N-sulfate D-glucuronosyl 5-epimerase, heparosan-glucuronate 5-epimerase, Heparosan-N-sulfate-D-glucuronosyl-5-epimerase, HG-5epi, HNSG-5epi, HS C5-epimerase, HS glucuronyl C5-epimerase, Hsepi, RED-C5-epimerase
ECTree
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Engineering
Engineering on EC 5.1.3.17 - heparosan-N-sulfate-glucuronate 5-epimerase
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H584A
site-directed mutagenesis, the mutant shows 80% reduced activity compared to wild-type
N585A
site-directed mutagenesis, the mutant shows 90% reduced activity compared to wild-type
R154A
site-directed mutagenesis, the mutant shows 60% reduced activity compared to wild-type
R156A
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
R396A
site-directed mutagenesis, the mutant shows 90% reduced activity compared to wild-type
R531A
site-directed mutagenesis, the mutant shows about 90% reduced activity compared to wild-type
R543A
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y149F
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y177F
site-directed mutagenesis, the mutant shows 40% reduced activity compared to wild-type
Y179F
site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type
Y468A
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y468F
site-directed mutagenesis, the mutant shows 75% reduced activity compared to wild-type
Y482F
site-directed mutagenesis, the mutant shows 20% reduced activity compared to wild-type
Y528A
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y528F
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y546A
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y546F
site-directed mutagenesis, the mutant shows over 90% reduced activity compared to wild-type
Y146A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y162A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y210A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
additional information
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construction of enzyme deficient strains ussing P-element transposase exscision mutagenesis, removal of the entire Hsepi coding sequence, generation of viable and fertile homozygous mutants for Hsepid12 and Hsepid13
additional information
high cell density fed-batch cultivation of recombinant Escherichia coli strains expressing 2-O-sulfotransferase and C5-epimerase at high level for the production of bioengineered heparin, method, overview. The first enzymatic step in this process uses heparan sulfate biosynthetic enzymes, 2-O-sulfotransferase (2-OST) and C5-epimerase (C5-epi), expressed as MBP-tagged proteins in Escherichia coli, to convert N-sulfo heparosan into an intermediate polysaccharide rich in -GlcNS(1->4)IdoA2S- sequences (where S is sulfo and IdoA is alpha-L-iduronic acid). This critical step in bioengineered heparin preparation relies on the use of recombinant arylsulfotransferase IV (AST-IV) to regenerate 3'-phospho adenosine-5'-phosphosulfate (PAPS) using p-nitrophenylsulfate as a sacrificial sulfur donor, one-pot reaction
additional information
generation of a soluble form of the enzyme HG-5epi by replacement of the transmembrane domain (N-terminal 33 amino acids) with the immunoglobulin Kappa signal sequence and a FLAG tag
additional information
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generation of a soluble form of the enzyme HG-5epi by replacement of the transmembrane domain (N-terminal 33 amino acids) with the immunoglobulin Kappa signal sequence and a FLAG tag
additional information
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generation of Glce null mutant mice, that show a strongly reduced size of the fetal spleen and a spectrum of defects in thymus and lymph node development ranging from dislocation to complete loss of the organ, overview. Transplantation of wild-type lymph nodes allows undisturbed lymphocyte maturation, phenotype, overview
additional information
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generation of Hsepi null mutant mice showing a lethal phenotype with selective organ defects but remarkably little effect on other organ systems, phenotype, overview. Heparan sulfate produced by enzyme-deficient MEF cells is devoid of L-iduronic acid residues, but shows up-regulated N- and 6-O-sulfation compared with wild-type, Hsepi-/- MEF cells proliferate and migrate similarly to wild-type cells. Restricted proliferation and migration of Hsepi mutant cells in response to FGF2 stimulation
additional information
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generation of Glce null mutant mice, that show a strongly reduced size of the fetal spleen and a spectrum of defects in thymus and lymph node development ranging from dislocation to complete loss of the organ, overview. Transplantation of wild-type lymph nodes allows undisturbed lymphocyte maturation, phenotype, overview
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