Information on EC 2.4.1.212 - hyaluronan synthase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.212
-
RECOMMENDED NAME
GeneOntology No.
hyaluronan synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] = UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
show the reaction diagram
-
-
-
-
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
SYSTEMATIC NAME
IUBMB Comments
Alternating UDP-alpha-N-acetyl-D-glucosamine:beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] 4-N-acetyl-beta-D-glucosaminyltransferase and UDP-alpha-D-glucuronate:N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] 3-beta-D-glucuronosyltransferase
The enzyme from Streptococcus Group A and Group C requires Mg2+. The enzyme adds GlcNAc to nascent hyaluronan when the non-reducing end is GlcA, but it adds GlcA when the non-reducing end is GlcNAc [3]. The enzyme is highly specific for UDP-GlcNAc and UDP-GlcA; no copolymerization is observed if either is replaced by UDP-Glc, UDP-Gal, UDP-GalNAc or UDP-GalA. Similar enzymes have been found in a variety of organisms.
CAS REGISTRY NUMBER
COMMENTARY hide
39346-43-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Mus musculus C57BL/6
-
-
-
Manually annotated by BRENDA team
Pasteurella multocida type A
type A
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Xenopus laevis protein DG42
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
regulation of hexosamine biosynthetic pathway, biosynthesis of hyaluronan and other glycoconjugates, and protein O-GlcNAcylation, overview
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
hyaluronic acid tetrasaccharide + UDP-alpha-D-glucuronate
?
show the reaction diagram
-
-
-
-
?
hyaluronic acid tetrasaccharide + UDP-alpha-N-acetyl-D-glucosamine
?
show the reaction diagram
-
-
-
-
?
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
show the reaction diagram
UDP-D-glucosamine + UDP-D-glucuronate
[beta-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + n UDP
show the reaction diagram
-
-
-
-
?
UDP-D-glucosamine + UDP-D-glucuronate
[beta-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
show the reaction diagram
-
-
-
-
?
UDP-D-glucuronate + chondroitin 4-sulfate trisaccharide
?
show the reaction diagram
-
3.6% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin 6-sulfate pentasaccharide
?
show the reaction diagram
-
61% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin 6-sulfate trisaccharide
?
show the reaction diagram
-
80% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + chondroitin sulfate
?
show the reaction diagram
-
12% of the activity with hyaluronan
-
-
?
UDP-D-glucuronate + unsulfated chondroitin
?
show the reaction diagram
-
54% of the activity with hyaluronan
-
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
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
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1-4)-beta-D-glucuronosyl-(1-3)-N-acetyl-beta-D-glucosaminyl-(1-4)-[nascent hyaluronan]
show the reaction diagram
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan]
show the reaction diagram
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
-
activates at 50 mM, slightly more effective than NaCl
NaCl
-
activates at 50 mM, slightly less effective than KCl
additional information
-
metal requirements of mutant enzymes, overview
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-deoxyglucose
-
-
4-methylesculetin
-
-
4-Methylumbelliferone
5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside
-
modulates aortic smooth muscle cell motility and adhesive properties through AMP-activated protein kinase, AMPK
AG825
ErbB2 inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation; ErbB2 inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation; ErbB2 inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation
HAS1 siRNA
-
-
-
iodoacetamide
mannose
inhibits HAS because it depletes UDP-GlcNAc in cells; inhibits HAS because it depletes UDP-GlcNAc in cells; inhibits HAS because it depletes UDP-GlcNAc in cells
metformin
-
modulates aortic smooth muscle cell motility and adhesive properties through AMP-activated protein kinase, AMPK
methylmethanethiosulfonate
N-ethylmaleimide
PEG
-
increasing MW increases the inhibitory effect on the enzyme, overview
Sodium arsenite
sodium salicylate
-
sodium salicylate is a potent suppressor of HAS1 activation
sulfhydryl reagents
tetramyristoyl cardiolipin
-
inactivating
thiazolidinedione
ERK inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation; ERK inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation; ERK inhibitor, blocks the heregulin-mediated HAS isozyme phosphorylation/activation
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
black rice hydrolized peptides
-
BRP from germinated Oryza sativa L. var. japonica
-
carbocyclic thromboxane A2
-
-
cardiolipin
-
required for full activity, activates up to 10fold, 14-18 molecules of cardiolipin are bound to one molecule of enzyme
dioleoyl phosphatidic acid
-
stimulates about 10fold
Epidermal growth factor
-
EGF
Epstein-Barr virus
-
-
-
ERK
heregulin-mediated HAS isozyme phosphorylation/activation; heregulin-mediated HAS isozyme phosphorylation/activation; heregulin-mediated HAS isozyme phosphorylation/activation
-
interleukin 1beta
-
activation of enzyme in plasma membrane fraction
-
Interleukin-1beta
O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
-
increases O-GlcNAcylation and HA synthesis
PDGF-BB
-
-
-
phorbol 12-myristate 13-acetate
-
activation of enzyme in plasma membrane fraction
pinane thromboxane A2
-
-
platelet derived growth factor BB
platelet derived growth factor-BB
-
PDGF-BB
-
platelet-derived growth factor BB
-
activation of enzyme in plasma membrane fraction
-
prostacyclin
-
-
prostaglandin E2
prostaglandin F2alpha
-
-
prostaglandin I2
synthetic single-stranded poly(A)
-
-
-
synthetic single-stranded poly(C)
-
-
-
synthetic viral RNA analog poly(I,C)
-
-
-
tetraoleoyl cardiolipin
TGFbeta1
-
-
-
TNFalpha
-
-
-
Transforming growth factor
-
TGF-1beta
-
transforming growth factor-beta
-
TGF-beta
-
transforming growth factor-beta1
-
TGF-beta1
-
tunicamycin
-
significant increase in HAS activity in the cytosolic membrane fraction after tunicamycin treatment
UVB irradiation
-
UVB increases HAS2 mRNA expression; UVB increases HAS3 mRNA expression
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.91
hyaluronic acid tetrasaccharide
-
25C, pH 7.5
0.014
UDP-alpha-D-glucuronate
-
25C, pH 7.5
0.66
UDP-alpha-N-acetyl-D-glucosamine
-
25C, pH 7.5
0.032 - 0.93
UDP-D-glucuronate
0.053 - 1.1
UDP-N-acetyl-D-glucosamine
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
120
UDP-D-glucuronate
Streptococcus dysgalactiae subsp. equisimilis
-
recombinant enzyme, pH 7.0, 30C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.5
ethylene glycol
-
recombinant enzyme, pH 7.0, 30C
3.3
glycerol
-
recombinant enzyme, pH 7.0, 30C
3.5
PEG 20000
-
recombinant enzyme, pH 7.0, 30C
1.2
Sucrose
-
recombinant enzyme, pH 7.0, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000000062
HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment, plus heregulin; HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment, plus heregulin; HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment, plus heregulin
0.0000000092
HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment; HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment; HAS activity, HAS1siRNA + HAS2siRNA + HAS3siRNA treatment
0.000000017
HAS activity, HAS2siRNA treatment; HAS activity, HAS2siRNA treatment; HAS activity, HAS2siRNA treatment
0.0000000175
HAS activity, HAS1siRNA treatment; HAS activity, HAS1siRNA treatment; HAS activity, HAS1siRNA treatment
0.0000000188
HAS activity, HAS3siRNA treatment; HAS activity, HAS3siRNA treatment; HAS activity, HAS3siRNA treatment
0.0000000222
HAS activity, AG825 treatment; HAS activity, AG825 treatment; HAS activity, AG825 treatment
0.0000000227
HAS activity, HAS2siRNA treatment, plus heregulin; HAS activity, HAS2siRNA treatment, plus heregulin; HAS activity, HAS2siRNA treatment, plus heregulin
0.000000023
HAS activity, thiazolidinedione treatment; HAS activity, thiazolidinedione treatment; HAS activity, thiazolidinedione treatment
0.0000000233
HAS activity, no treatment, control; HAS activity, no treatment, control; HAS activity, no treatment, control
0.0000000237
HAS activity, scrambled sequence treatment, control; HAS activity, scrambled sequence treatment, control; HAS activity, scrambled sequence treatment, control
0.0000000248
HAS activity, HAS1siRNA treatment, plus heregulin; HAS activity, HAS1siRNA treatment, plus heregulin; HAS activity, HAS1siRNA treatment, plus heregulin
0.000000025
HAS activity, HAS3siRNA treatment, plus heregulin; HAS activity, HAS3siRNA treatment, plus heregulin; HAS activity, HAS3siRNA treatment, plus heregulin
0.0000000257
HAS activity, AG825 treatment, plus heregulin; HAS activity, AG825 treatment, plus heregulin; HAS activity, AG825 treatment, plus heregulin
0.0000000263
HAS activity, thiazolidinedione treatment, plus heregulin; HAS activity, thiazolidinedione treatment, plus heregulin; HAS activity, thiazolidinedione treatment, plus heregulin
0.0000000583
HAS activity, scrambled sequence treatment, control, plus heregulin; HAS activity, scrambled sequence treatment, control, plus heregulin; HAS activity, scrambled sequence treatment, control, plus heregulin
0.0000000627
HAS activity, no treatment, control, plus heregulin; HAS activity, no treatment, control, plus heregulin; HAS activity, no treatment, control, plus heregulin
13.4
-
mutant C226A/C262A, pH 7.0, 30C
14
-
deletion mutant DELTA3-C281, pH 7.0, 30C
16.4
-
mutant C226A, pH 7.0, 30C
16.5
-
mutant C226S, pH 7.0, 30C
21.2
-
mutant C281S, pH 7.0, 30C
22.9
-
deletion mutant C-Null, pH 7.0, 30C
25.3
-
mutant C226A/C281A, pH 7.0, 30C
27
-
deletion mutant DELTA3-C367, pH 7.0, 30C
29.1
-
mutant C262A, pH 7.0, 30C
29.5
-
mutant C367S, pH 7.0, 30C
30.2
-
mutant C262A/C281A, pH 7.0, 30C
31.4
-
mutant C262S, pH 7.0, 30C
31.6
-
deletion mutant DELTA3-C262, pH 7.0, 30C
33.7
-
mutant C281A, pH 7.0, 30C
37
-
wild-type enzyme, pH 7.0, 30C
37.8
-
mutant C226A/C367A, pH 7.0, 30C
38.6
-
deletion mutant DELTA3-C226, pH 7.0, 30C
39.5
-
mutant C281A/C367A, pH 7.0, 30C
46.7
-
mutant C367A, pH 7.0, 30C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
assay at
7.6 - 8.1
-
-
9 - 10
-
recombinant enzyme, pH-profile
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 11.5
-
recombinant enzyme, pH-profile
7 - 8.4
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
fibroblast, expression of isozymes HAS1 and HAS2 is increased after oncogenic malignant transformation with v-sre and/or v-fos, while only the expression of isozyme HAS2 is increased by transformation with v-HA-ras; fibroblast, oncogenic malignant transformation with v-sre and/or v-fos, and v-HA-ras
Manually annotated by BRENDA team
expression of HAS1 splice variants is absent from B cells of healthy donors and in multiple myeloma and monoclonal gammopathy of undetermined significance (MGUS) is restricted to the B-cell compartment
Manually annotated by BRENDA team
-
skin dermal fibroblast cell
Manually annotated by BRENDA team
at embryonic day 7.5, Has1 is expressed throughout the gastrulating embryo. After ambryonic day 8.5, Has1 expression disappears; at embryonic day 7.5, Has2 is expressed throughout the gastrulating embryo. After ambryonic day 8.5, Has2 continues to be strongly, albeit transiently, expressed in numerous tissues, including the branchial arches and craniofacial structures such as the palatal shelves and lens pit. Has2 is also expressed during cardiac, skeletal, and tail development; Has3 transcripts are first detected at embryonic day 10.5 in the maxillary and mandibular components of the first branchial arch. Has3 expression in the developing teeth, vibrissae hair follicles, nasal cavity, and inner ear complements the expression pattern of Has2
Manually annotated by BRENDA team
-
vitreous body
Manually annotated by BRENDA team
-
cell line
Manually annotated by BRENDA team
tumorigenic cell line, 3-5fold higher hyaluronan accumulation than in the parental cell line 3Y1; tumorigenic cell line, 3-5fold higher hyaluronan accumulation than in the parental cell line 3Y1
Manually annotated by BRENDA team
-
HAS1 expression is shown in 55.7% of SIRPA-positive macrophages in stage III follicles
Manually annotated by BRENDA team
-
mouse epithelial cell line
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
expression of isoform HAS1 in theca cells of healthy and early atretic follicles of stage I and stage II and in progressing atretic stage III follicles
Manually annotated by BRENDA team
-
serous epithelial ovarian tumor
Manually annotated by BRENDA team
-
serous epithelial ovarian tumor
Manually annotated by BRENDA team
-
platelet-derived growth factor BB exerts dominant influence on HAS2 isoform expression by osteoblasts
Manually annotated by BRENDA team
-
type DPK-SKDF-H
Manually annotated by BRENDA team
metastatic cell line, 3-5fold higher hyaluronan accumulation than in the parental cell line 3Y1; metastatic cell line, 3-5fold higher hyaluronan accumulation than in the parental cell line 3Y1
Manually annotated by BRENDA team
-
expression of isoform HAS1 in theca cells of healthy and early atretic follicles of stage I and stage II and in progressing atretic stage III follicles
Manually annotated by BRENDA team
-
35 endometrial tissue biopsies from 35 patients, including proliferative and secretory endometrium, post-menopausal proliferative endometrium, complex atypical hyperplasia, grade 1 and grade 2 + 3 endometrioid adenocarcinomas. Immunoreactivity of all HAS proteins is increased in the cancer epithelium
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
48000
-
northern blot
52000
-
SDS-PAGE
60000
determined by SDS-PAGE and Western Blot analysis; determined by SDS-PAGE and Western Blot analysis
62000
determined by SDS-PAGE and Western Blot analysis
66000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 49000, recombinant enzyme, SDS-PAGE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
HAS activity can be modulated by post-translational modification, such as phosphorylation and N-glycosylation
phosphoprotein
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
1 h, at 30C, loss of 74-90% activity
658093
11.5
-
1 h, at 30C, loss of 40-50% activity
658093
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
the enzyme is temperature labile, but is stabilized by substrate and cardiolipin
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
HAS2 can be O-GlcNAcylated on serine 221, which strongly increases its activity and its stability to half-life of over 5 h versus 17 min without O-GlcNAcylation
-
substrate and cardiolipin stabilize the enzyme
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, Na-phosphate buffer, 10% glycerol, 96 h, 18%
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
on a Ni2+-nitrilotriacetic acid resin
-
partial
-
recombinant enzyme from Escherichia coli, native enzyme to homogeneity
-
recombinant enzyme from membranes of Escherichia coli, to homogeneity
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
-
recombinant His6-tagged enzyme from Escherichia ccoli strain C43 by solubilization with detergent LysoFosCholine Ether-14, nickel affinity chromatography, and gel filtration
-
recombinant isozymes by immunoaffinity chromatography
recombinant synthesis of hyaluronan is carried out with Agrobacterium sp. strain ATCC 31749, hyaluronan is primarly found in the culture medium
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a plasmid encoding FLAG epitope-tagged HAS3 for transfection of human prostate tumor cells is constructed, Tet-inducible 22Rv1 cells are generated
-
DNA and amino acid sequence determination and analysis, chromosome mapping of isozymes HAS1-3, genetic organization
-
ectopic expression of Flag- and 6myc-HAS2 in COS-1 cells as homodimers, co-expression with Flag-HAS3 leads to formation of heterodimers
-
expression as a soluble active protein comprising residues 1-703
-
expression in COS-1 cells and rat 3Y1 fibroblasts
-
expression in COS-7 cells
expression in Escherichia coli
expression in Saccharomyces cerevisiae and Escherichia coli
-
expression in yeast
-
expression of both hyaluronan synthase and UDP-glucose-6-dehydrogenase in Lactococcus lactis
-
expression of C-terminally His6-tagged Se-HAS in Escherichia coli strain C43
-
expression of His-tagged enzyme in Escherichia coli
-
expression of His-tagged wild-type and mutant enzymes in Escherichia coli
-
expression of isozyme HAS2-MBP-fusion protein in Escherichia coli strain JM109
-
expression of mutant enzymes in Escherichia coli. SeHAS(E327Q) and seHAS(E327K) are expressed at low levels, whereas seHAS(E327D) and the Lys48 mutants are expressed well
-
expression of wild-type and mutant enzymes as His-tagged proteins in Escherichia coli
-
expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain BJ5461
-
expression of wild-type and mutants enzymes in Escherichia coli
-
expression of wild-type FLAG-tagged human HAS1, HAS2, or HAS3, and of HAS T110A mutant enzyme in COS-7 cells
-
functional expression of isozymes HAS2 and HAS3 in rat epidermal keratinocytes as N-terminally GFP-tagged protein, recombinant isozymes GFP-HAS2 and GFP-HAS3 travel through endoplasmic reticulum, Golgi, plasma membrne, and endocytic vesicles, expression of inactive GFP-tagged HAS3 D216A mutant and of GFP-tagged HAS3-deletion mutants in keratinocytes
-
gene HAS1, expression analysis and recombinant expression in COS-1 cells; gene HAS2, expression analysis and recombinant expression in COS-1 cells; gene HAS3, expression analysis and recombinant expression in COS-1 cells
genes HAS1, HAS2 and HAS3, quantitative expression analysis in benign and malign endometrial tissue
-
HAS1 DNA and amino acid sequence determination and analysis, oncogenic malignant transformation of 3Y1 fibroblasts with v-sre and/or v-fos, or v-HA-ras; HAS2 DNA and amino acid sequence determination and analysis, oncogenic malignant transformation of 3Y1 fibroblasts with v-sre and/or v-fos, or v-HA-ras
Homo sapiens hyaluronan synthase 1 is cloned into the plasmid pFLAG-CMV2, for in vitro translation the pF3A WG(BYDV) Flexi vector is used
-
into a pCR2.1 vector for sequencing; into a pCR2.1 vector for sequencing; into a pCR2.1 vector for sequencing
overexpression in Escherichia coli, enzyme cannot be expressed as a soluble active protein
-
overexpression of GFP-tagged HAS-1 in the wounds of lentiviral-HAS-1-treated mice
-
retroviral transduction system is used to overexpress the three murine hyaluronan synthase enzymes in arterial smooth muscle cells. Overexpression of hyaluronan synthases alters vascular smooth muscle cell phenotype and promotes monocyte adhesion
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stable expression of N-terminally Myc-tagged human HAS2 in NIH3T3 cells membranes, the molecular mechanism that underlies the rapid c-Myc-HAS2 turnover involves the 26 S proteasome, overview
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Streptococcus equisimilis hyaluronan synthase is cloned into the plasmid pKK223-3 for expression in Escherichia coli SURE cells
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the coding sequence of HAS1 is cloned into the vector pCR3.1 for transfection of fibroblasts
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the Escherichia coli expression vector pQE80L and the broad host range cloning vector pBBR122 are used
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the human HAS2 expression plasmid is prepared by inserting its coding sequence into the vector pcDNA 3.1/CT-GFP-TOPO
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
basic fibroblast growth factor bFGF increases HA-synthase-1 and -2 expression and enhances high molecular weight hyaluronan deposition in the pericellular matrix
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both tumor necrosis factor TNFalpha and interferon INFgamma significantly induce HAS3 expression; both tumor necrosis factor TNFalpha and transforming growth factor 1beta significantly increase HAS1 expression and protein synthesis. Exposure to reactive oxygen species results in increased gene expression and protein formation of HAS1; both tumor necrosis factor TNFalpha and transforming growth factor 1beta significantly increase HAS2 expression and protein synthesis. Exposure to reactive oxygen species results in increased gene expression and protein formation of HAS2
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chondroitin sulfate increases hyaluronan production by osteoarthritic fibroblast-like synoviocytes through up-regulation of the expression of HAS1 and HAS2 associated with activation of ERK-1/2, p38, and Akt, although to a lesser extent. Both p38 and Akt are involved in chondroitin sulfate-induced hyaluronan accumulation. IL-1 increases hyaluronan production and levels of mRNA for HAS1, HAS2, and HAS3. Chondroitin sulfate enhances the IL-1induced level of HAS2 mRNA and reduces the level of HAS3 mRNA. IL-1induced activation of p38 and JNK is slightly decreased by chondroitin sulfate, whereas that of ERK-1/2 and Akt is enhanced. More high molecular weight hyaluronan is found in chondroitin sulfate plus IL-1treated fibroblast-like synoviocytes than in fibroblast-like synoviocytes treated with IL-1 alone; chondroitin sulfate increases hyaluronan production by osteoarthritic fibroblast-like synoviocytes through up-regulation of the expression of HAS1 and HAS2 associated with activation of ERK-1/2, p38, and Akt, although to a lesser extent. Both p38 and Akt are involved in chondroitin sulfate-induced hyaluronan accumulation. IL-1 increases hyaluronan production and levels of mRNA for HAS1, HAS2, and HAS3. Chondroitin sulfate enhances the IL-1induced level of HAS2 mRNA and reduces the level of HAS3 mRNA. IL-1induced activation of p38 and JNK is slightly decreased by chondroitin sulfate, whereas that of ERK-1/2 and Akt is enhanced. More high molecular weight hyaluronan is found in chondroitin sulfate plus IL-1treated fibroblast-like synoviocytes than in fibroblast-like synoviocytes treated with IL-1 alone; chondroitin sulfate increases hyaluronan production by osteoarthritic fibroblast-like synoviocytes through up-regulation of the expression of HAS1 and HAS2 associated with activation of ERK-1/2, p38, and Akt, although to a lesser extent. Both p38 and Akt are involved in chondroitin sulfate-induced hyaluronan accumulation. IL-1 increases hyaluronan production and levels of mRNA for HAS1, HAS2, and HAS3. Chondroitin sulfate enhances the IL-1induced level of HAS2 mRNA and reduces the level of HAS3 mRNA. IL-1induced activation of p38 and JNK is slightly decreased by chondroitin sulfate, whereas that of ERK-1/2 and Akt is enhanced. More high molecular weight hyaluronan is found in chondroitin sulfate plus IL-1treated fibroblast-like synoviocytes than in fibroblast-like synoviocytes treated with IL-1 alone
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comparison of full-length HAS1 isoform and its splice variants Va, Vb, and Vc. When co-expressed, the properties of HAS1 variants are dominant over those of full length HAS1. Full length HAS1 appears to be diffusely expressed in the cell, but HAS1 variants are concentrated in the cytoplasm and/or Golgi apparatus. HAS1 variants synthesize detectable de novo hyaluronan intracellularly. Each of the HAS1 variants is able to relocalize full length HAS1 protein from diffuse cytoskeleton-anchored locations to deeper cytoplasmic spaces. The HAS1-variants-mediated relocalization occurs through strong molecular interactions, which also serve to protect full length HAS1 from its otherwise high turnover kinetics
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during atresia, HAS1 mRNA and protein expression is markedly up-regulated inovary. HAS2 and HAS3 mRNA expression levels are low and very low to undetectable, respectively
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HAS2 is the major isoenzyme in MCF-7 cells. The mRNA expression is lowered by 4-methylumbelliferone by 81% in MCF-7 cells, and by 88% in A2058 cells. Both HAS substrate and HAS2 and/or HAS3 mRNA are targeted by 4-methylumbelliferone. The reduction of hyaluronan caused by 4-methylumbelliferone is associated with a significant inhibition of cell migration, proliferation and invasion; in MDA-MB-361, A2058 and SKOV-3 cells, treatment with 4-methylumbelliferone decreases HAS3 mRNA by 84-60%. The reduction of hyaluronan caused by 4-methylumbelliferone is associated with a significant inhibition of cell migration, proliferation and invasion
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isoform HAS2 is a primary target of the cAMP activator forskolin and the nuclear hormone alltrans-retinoic acid. Forskolin and all-trans-retinoic acid modulate the formation of complexes between transcription factor CREB1 and retinoic acid receptor with various co-regulators at the predicted sites.Mediator MED1 and co-repressor nuclear receptor co-repressor NCoR1 are central for the all-trans-retinoic acid induction of the HAS2 gene and CREB-binding protein CBP dominates its forskolin response
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reduced HAS 2 gene expression and increased excreted urinary hyaluronidase activity during dehydration contribute to the reduced amount of hyaluronan and to antidiuretic response
transforming grwoth factor 1beta significantly inhibits HAS3 expression and protein formation
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S221A
-
site-directed mutation of the O-GlcNAcylable Ser-221 to alanine generated an enzyme with a calculated t1/2 of about 70 min
T110A
-
site-directed mutagenesis of the phosphorylation site residue, the mutant is not inhibited by AMP-activated protein kinase
D216A
-
site-directed mutagenesis, isozyme HAS3, inactive mutant
K190R
-
site-directed mutagenesis, inactive mutant, K190R-mutated HAS2 forms dimers with wild-type HAS2 and quenches the activity of wild-type HAS2
D196N
-
mutants possess UDP-D-glucuronate-transferase activity
D247E
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D247K
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D247N
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249E
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249K
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D249N
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
D370E
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low hyaluronan synthase activity
D370K
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
D370N
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
D477K
-
mutants possess UDP-N-acetyl-D-glucosamine-transferase activity
D527E
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D527K
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D527N
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
D529E
-
site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and low hyaluronan synthase activity
D529K
-
site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and very low hyaluronan synthase activity
D529N
-
site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity
E369D
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
E369H
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
E369Q
-
site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity
D247E
Pasteurella multocida type A
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
-
D247K
Pasteurella multocida type A
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
-
D247N
Pasteurella multocida type A
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
-
D249N
Pasteurella multocida type A
-
site-directed mutagenesis, mutant possesses only GlcUA-transferase activity
-
C226A/C262A
C226A/C262A/C367A
-
site-directed mutagenesis, 1.4% remaining activity and altered kinetic constants compared to the wild-type enzyme
C226A/C281A
C226A/C367A
C262A/C281A
C262A/C367A
C281A/C367A
E327D
-
the specific enzyme activity relative to wild type enzyme is 38%. Mutant enzyme synthesizes hyaluronan of smaller weight-average molar mass than wild-type enzyme
E327K
-
the specific enzyme activity relative to wild type enzyme is 0.16%. Mutant enzyme synthesizes hyaluronan of smaller weight-average molar mass than wild-type enzyme
E327K/K48E
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the specific enzyme activity near wild-type level. Mutant enzyme synthesizes hyaluronan of smaller weight-average molar mass than wild-type enzymel
E327Q
-
the specific enzyme activity relative to wild type enzyme is 26%. Mutant enzyme synthesizes hyaluronan of smaller weight-average molar mass than wild-type enzyme
K48F
-
site-directed mutagenesis, alteration of K48 within membrane domain 2 causes decreased activity and HA product size
K48R
-
the specific enzyme activity relative to wild type enzyme is 17%. Mutant enzyme synthesizes hyaluronan of smaller weight-average molar mass than wild-type enzyme
C117F
-
site-directed mutagenesis, no expression in yeast possible
C117L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
C117S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme
C210S
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
C239S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme
C239S/C337S
-
site-directed mutagenesis, reduced recombinant expression level, activity is similar to the wild-type enzyme
C298F
-
site-directed mutagenesis, poor recombinant expression level, highly reduced activity compared to the wild-type enzyme
C298L
-
site-directed mutagenesis, poor recombinant expression level, highly reduced activity compared to the wild-type enzyme
C298S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme
C304S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme
C304S/C337S
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
C307S
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
C307S/C337S
-
site-directed mutagenesis, reduced recombinant expression level, inactive mutant
C337S
-
site-directed mutagenesis, increased Km for UDP-N-acetylglucosamine compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
-
optimization of the recombinant enzyme expression in Escherichia coli for large scale production of hyaluronan polymers for usage in basic studies, and for biotechnological creation of functional carbohydrates in medical purposes, engineering of produced product chain length
medicine
synthesis
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