Information on EC 3.1.6.4 - N-acetylgalactosamine-6-sulfatase

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

EC NUMBER
COMMENTARY hide
3.1.6.4
-
RECOMMENDED NAME
GeneOntology No.
N-acetylgalactosamine-6-sulfatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of the 6-sulfate groups of the N-acetyl-D-galactosamine 6-sulfate units of chondroitin sulfate and of the D-galactose 6-sulfate units of keratan sulfate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of sulfuric ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycosaminoglycan degradation
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
N-acetyl-D-galactosamine-6-sulfate 6-sulfohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
59299-00-2
-
9025-60-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain IFO-13310
-
-
Manually annotated by BRENDA team
strain IFO-13310
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Japonica
-
-
Manually annotated by BRENDA team
Japonica
-
-
Manually annotated by BRENDA team
isolated from dog otic secretion or skin
-
-
Manually annotated by BRENDA team
precursor
UniProt
Manually annotated by BRENDA team
no activity in Streptococcus anginosus
-
-
-
Manually annotated by BRENDA team
isolated from humans, a penguin, an armadillo and dog food
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
modification of expression of the enzyme regulates the content of chondroitin sulfate
physiological function
in post-natal ventral rat prostate, a distinct and reciprocal localization of arylsulfatase B and N-acetylgalactosamine-6-sulfatase is seen, with arylsulfatase B predominant in the stroma and N-acetylgalactosamine-6-sulfatase predominant in the epithelium. Estrogen treatment does not inhibit the increase in GALNS activity between days 5 and 30
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl beta-D-galactoside 6-sulfate + H2O
4-methylumbelliferyl beta-D-galactoside + sulfate
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl sulfate + H2O
4-methylumbelliferone + sulfate
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl-beta-D-galactopyranoside-6-sulfate + H2O
4-methylumbelliferyl-beta-D-galactopyranoside + sulfate
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl-beta-D-galactose-6-sulfate + H2O
4-methylumbelliferyl-beta-D-galactose + sulfate
show the reaction diagram
-
-
-
-
?
beta-N-acetyl-D-galactosamine-6-sulfate-(1-4)-beta-D-glucuronic acid-(1-3)-N-acetyl-D-galactosaminitol 6-sulfate + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
chondroitin sulfate + H2O
chondroitin + sulfate
show the reaction diagram
chondroitin-6-sulfate + H2O
chondroitin + sulfate
show the reaction diagram
Gal(6-SO4)beta(1-4)-GlcAc(6-SO4)beta(1-4)-D-galactitol + H2O
sulfate + ?
show the reaction diagram
-
-
-
?
galactose 6-sulfate + H2O
galactose + sulfate
show the reaction diagram
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)2 + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)3 + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)4 + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
GalNAc-4-SO4-GlcUA-GalNAc-4-SO4 + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
keratan sulfate + H2O
keratan + sulfate
show the reaction diagram
N-acetyl-D-glucosamine 6-sulfate units + H2O
N-acetyl-D-glucosamine units + sulfate
show the reaction diagram
-
in chondroitin sulfate or keratan sulfate, decreased enzymatic activity in fasting rats
-
?
N-acetylgalactosamine 6-sulfate + H2O
N-acetylgalactosamine + sulfate
show the reaction diagram
N-acetylgalactosamine 6-sulfate-anhydromannitol + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
N-acetylgalactosamine 6-sulfate-beta(1-4)glucuronic acid-beta(1-3)-N-acetylgalactosaminitol 6-sulfate + H2O
sulfate + ?
show the reaction diagram
-
sulfate is hydrolyzed only from the non-reducing terminal
-
-
?
O-(beta-D-6-sulfo-2-acetamido-2-deoxygalactosyl)-(1-4)-O-beta-D-glucuronosyl-(1-3)-O-beta-D-6-sulfo-2-acetamido-2-deoxy-galactitol + H2O
sulfate + ?
show the reaction diagram
-
-
-
-
?
O-(beta-D-sulfogalactosyl)-(1-4)-1,5-anhydro-D-mannitol 6-sulfate + H2O
?
show the reaction diagram
-
-
-
-
?
sulfated tetrasaccharide + H2O
?
show the reaction diagram
-
sulfated tetrasaccharide obtained by digestion of purified chondroitin 6-sulfate with resticular hyaluronidase
-
-
?
tetrasaccharide trisulfate + H2O
sulfate + N-acetylglucosamine + galactose
show the reaction diagram
trisaccharide disulfate + H2O
sulfate + ?
show the reaction diagram
UDP-N-acetylgalactosamine 6-sulfate + H2O
sulfate + UDP-N-acetylgalactosamine
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
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-Sulfated heptasaccharides
-
strong
-
6-Sulfated pentasaccharides
-
strong
-
chondroitin 4-sulfate
-
slight
chondroitin 6-sulfate
-
competitive
Chondroitin 6-sulfate tetrasaccharide
-
slight
-
dermatan sulfate
-
slight
Fe3+
-
-
heparan sulfate
-
competitive
heparin
Hg2+
-
-
hyaluronic acid
-
slight
Keratan sulfate
-
competitive
Mg2+
-
-
Mn2+
-
-
Na2HPO4
-
-
NaH2PO4
-
-
NO3-
-
-
S2O32-
-
-
SO32-
-
-
Zn2+
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
activates
dithiothreitol
-
activates
EDTA
-
activates
glutathione
-
activates
NEM
-
activates
additional information
-
cotransfection with sulfatase modifying factor 1 shows an increment up to 2.6fold in GALNS activity, cells cotransfected with elongation factor 1alpha show GALNS activity continued to increase until day 8 postransfection
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.4
4-Methylumbelliferyl sulfate
-
recombinant enzyme
0.008
galactosamine 6-sulfate
-
-
0.013
Galactose 6-sulfate
-
-
0.03
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)2
-
-
-
0.049
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)3
-
-
-
0.064
GalNAc-4-SO4-(GlcUA-GalNAc-4-SO4)4
-
-
-
0.027
GalNAc-4-SO4-GlcUA-GalNAc-4-SO4
-
-
-
0.12
N-acetylgalactosamine 6-sulfate-beta(1-4)glucuronic acid-beta(1-3)-N-acetylgalactosaminitol 6-sulfate
-
-
-
0.012 - 0.015
O-(beta-D-6-sulfo-2-acetamido-2-deoxygalactosyl)-(1-4)-O-beta-D-glucuronosyl-(1-3)-O-beta-D-6-sulfo-2-acetamido-2-deoxy-galactitol
0.05 - 0.096
O-(beta-D-sulfogalactosyl)-(1-4)-1,5-anhydro-D-mannitol 6-sulfate
0.0125
oligosaccharide substrate derived from chondroitin 6-sulfate
-
-
-
0.05
oligosaccharide substrates derived from keratan sulfate
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-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000653
-
enzyme from fibroblasts
0.00073
-
enzyme from placenta
1.11
-
-
2.13
-
-
2.68
-
-
additional information
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-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 4
3.8 - 4
-
hydrolysis of N-acetylgalactosamine 6-sulfate or galactose 6-sulfate
4
-
recombinant enzyme
4.2
-
and a second optimum at pH 5.2, 0.1 M sodium acetate buffer
4.4
-
and a second optimum at pH 4.9, 0.02 M sodium acetate buffer
4.9
-
and a second optimum at pH 4.4, 0.02 M sodium acetate buffer
5.2
-
and a second optimum at pH 4.2, 0.1 M sodium acetate buffer
6.5
-
release of sulfate from galactose 6-sulfate and N-acetylgalactosamine 6-sulfate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.3 - 4.2
-
pH 3.0: about 45% of maximal activity, pH 4.2: about 55% of maximal activity, hydrolysis of N-acetylgalactosamine 6-sulfate, 0.02 M acetate buffer
5 - 6
-
purified recombinant enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
in post-natal ventral rat prostate, a distinct and reciprocal localization of arylsulfatase B and N-acetylgalactosamine-6-sulfatase is seen, with arylsulfatase B predominant in the stroma and N-acetylgalactosamine-6-sulfatase predominant in the epithelium. Estrogen treatment does not inhibit the increase in GALNS activity between days 5 and 30
Manually annotated by BRENDA team
no detectable enzyme amount in Mucopolysaccheridosis type IVA patients
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52000 - 53000
-
N204K mutant, precursor protein lacking one carbohydrate chain, SDS-PAGE
55000
-
precursor protein, SDS-PAGE
57000
-
x * 19000, x * 39000, x * 57000, the active enzyme contains either the 57000 Da polypeptide or disulfide-linked 39000 Da and 19000 Da polypeptides
58000
-
recombinant precursor enzyme, gel filtration
62000
-
gel filtration
78000
-
x * 78000, SDS-PAGE
85000
-
x * 85000, SDS-PAGE
90000
-
gel filtration
100000
-
gel filtration
120000
additional information
-
in normal cells N-acetylgalactosamine-6-sulfate sulfatase is in a 1.27 MDa complex with three other lysosomal hydrolases: beta-galactosidase, alpha-neuraminidase, and cathepsin A
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 60000
homodimer
-
2 * 60000 Da
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
modeling of structure and molecular docking against galactose-6-sulfate, N-acetylgalactosamine-6-sulfate, keratan sulfate, chondroitin-6-sulfate, and the artificial substrate 4-methylumbelliferyl-beta-D-galactopyranoside-6-sulfate. Amino acids involved in ligand interaction correlate with those observed in other human sulfatases, and mutations within the active cavity reduce affinity of all evaluated ligands
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
30 min, 50% loss of actuvity
55
-
10 min, 50% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freezing and thawing five times do not decrease the enzyme activity
-
freezing to -20C and immediate thawing decreases the activity by 20%
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 2 months, 5% loss of activity
-
4C, 2 months, 20% loss of activity
-
4C, 25 mM sodium acetate, 1 mM beta-glycerophosphate buffer, pH 5.5, several months, no significant loss of acitivity
-
4C, stable for 10 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant precursor enzyme
-
two-step affinity chromatography, purified 1317-fold, 71% yield
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in deficient fibroblasts
-
expression in Chinese hamster ovary cells
-
expression in Escherichiacoli BL21
-
expression in MCF-7 cells. Following overexpression of the enzyme, total sulfated glycosaminoglycans, chondroitin 4-sulfate, and chondroitin sulfates decline significantly
-
overexpression in chinese hamster ovary cells
-
recombinant enzyme expression in Escherichia coli strain BL21 in both soluble and inclusion bodies fractions, rGALNS amounts in inclusion bodies fraction are up to 17fold higher than those observed in the soluble fraction, method optimization, effect of aeration and agitation at bench scale, detailed overview
-
the GALNS gene is located on chromosome 16q24.3, DNA and amino acid sequence determination and analysis of wild-type and mutant genes
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme activity but not protein content declines after lambda-carrageenan exposure (0.001 mg/l for 48 h)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A291D
-
mutation associated with attenuated phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, , in wild-type, A291 has a hydrogen bond with K310
A291T
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking. In wild-type, A291 has a hydrogen bond with K310. Mutation A291T produces a new hydrogen bond with G301
C79S
-
no enzymatic activity
C79T
-
no enzymatic activity
C79Y
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
F97V
-
10fold decrease in enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
G155R
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
G168R
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
G290S
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
G309R
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
G389del
-
mutation leads to mucopolysaccharidosis IVA
G929del
-
mutation leads to mucopolysaccharidosis IVA
H142R
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
H166Q
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
H236D
-
mutation associated with an attenuated phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, residue is involved in the ligand-enzyme interaction
I113F
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
K310N
-
mutation associated with attenuated phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, , residue is involved in enzyme-ligand interaction
L67M
-
N-acetylgalactosamine-6-sulfatase mutation of a mucopolysaccharidosis type IV tunisian patient
M318R
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
N204K
-
decreased enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
N289S
-
mutation associated with an attenuated phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, , residue is involved in the ligand-enzyme interaction
P125L
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
P151L
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
P179S
-
mutation leads to mucopolysaccharidosis IVA
P357L
-
mutation leads to mucopolysaccharidosis IVA
Q473X
-
mutation leads to mucopolysaccharidosis IVA
R295Q
-
decreased enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
R386C
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
R94C
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
R94G
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
S162F
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
S80L
-
mutation associated with a severe phenotype of lysosomal storage disease Mucopolysaccharidosis IV A, modeling of energy minimization and affinity energy after docking
T312S
-
decreased enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
T763del
-
mutation leads to mucopolysaccharidosis IVA
V138A
-
no enzymatic activity, no mature polypeptide chain and precursor polypeptide is faintly visible
C76S
active site mutation, results in inactive enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
synthesis
-
production and characterization of an active recombinant N-acetylgalactosamine-6-sulfate sulfatase in Escherichia coli BL21. When native signal peptide is present, higher enzyme activity levels are observed in both soluble and inclusion bodies fractions, and signal peptide removal has a significant impact on enzyme activation. Enzyme activity in the culture media is only detected when signal peptide is presented and the culture is carried out under semi-continuous mode