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Information on EC 3.1.6.8 - cerebroside-sulfatase and Organism(s) Homo sapiens
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3.1.6.8 cerebroside-sulfataseIUBMB Comments
Hydrolyses galactose-3-sulfate residues in a number of lipids. Also hydrolyses ascorbate 2-sulfate and many phenol sulfates.
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Word Map
- 3.1.6.8
-
metachromatic
- leukodystrophy
- sulfatide
- medicine
- lysosomal
- asa
-
demyelinate
-
sural
-
pseudodeficiency
- nitrocatechol
-
sulfatases
- molecular biology
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Synonyms
arylsulfatase-a, cerebroside sulfatase, cerebroside sulfate sulfatase, cerebroside-sulfatase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
ARSA
arylsulfatase A
ASA
cerebroside sulfatase
cerebroside sulfate sulfatase
-
-
-
-
Cerebroside-sulfatase
-
-
-
-
sulfatase, cerebroside
-
-
-
-
arylsulfatase A
-
-
-
-
arylsulfatase A
-
-
ASA
-
-
-
-
cerebroside sulfatase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a cerebroside 3-sulfate + H2O = a cerebroside + sulfate
hydrolyses galactose-3-sulfate residues in a number of lipids, also hydrolyses ascorbate 2-sulfate and many phenol sulfates, enzymes catalyze in addition to reaction of EC 3.1.6.8 also arylsulfatase reaction EC 3.1.6.1
-
a cerebroside 3-sulfate + H2O = a cerebroside + sulfate
A detailed mechanism for sulfate ester cleavage is proposed, involving an aldehyde hydrate as the functional group.
a cerebroside 3-sulfate + H2O = a cerebroside + sulfate
a detailed mechanism for sulfate ester cleavage is proposed, involving an aldehyde hydrate as the functional group
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of sulfuric ester
-
-
-
-
PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
cerebroside-3-sulfate 3-sulfohydrolase
Hydrolyses galactose-3-sulfate residues in a number of lipids. Also hydrolyses ascorbate 2-sulfate and many phenol sulfates.
CAS REGISTRY NUMBER
COMMENTARY
9068-68-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
1-O-alkyl-2-O-acyl-3-(beta-3'-sulfogalactosyl)glycerol + H2O
1-O-alkyl-2-O-acyl-3-O-(beta-galactosyl)glycerol + sulfate
1-O-alkyl-2-O-acyl-3-O-(beta-D-galactopyranoside-3'-sulfate)-glycerol + H2O
1-O-alkyl-2-O-acyl-3-O-beta-D-galactopyranoside-glycerol + sulfate
-
-
-
-
?
cerebroside 3-sulphate + H2O
cerebroside + sulfate
the enzyme catalyzes the first step in the degradation of the glycolipid cerebroside sulfate
-
?
lysoseminolipid sulfate + H2O
lysoseminolipid + sulfate
-
deacylated seminolipid
-
-
?
methylumbelliferyl sulfate + H2O
methylumbelliferone + sulfate
-
-
-
-
?
N-acyl-1-O-(beta-3-sulfogalactosyl)sphingosine + H2O
(N-acyl-1-O-galactosyl) sphingosine + sulfate
-
sulfosphingolipids
-
-
?
N-lissamine rhodaminyl-(12-aminododecanoyl)cerebroside 3-sulfate + H2O
?
-
-
-
-
?
nitrocatechol sulfate + H2O
nitrocatechol + sulfate
-
-
-
-
?
1-O-alkyl-2-O-acyl-3-(beta-3'-sulfogalactosyl)glycerol + H2O
1-O-alkyl-2-O-acyl-3-O-(beta-galactosyl)glycerol + sulfate
-
-
-
-
?
1-O-alkyl-2-O-acyl-3-(beta-3'-sulfogalactosyl)glycerol + H2O
1-O-alkyl-2-O-acyl-3-O-(beta-galactosyl)glycerol + sulfate
-
seminolipid, sulfoglycerolipids
-
-
?
4-nitrocatechol sulfate + H2O
4-nitrocatechol + sulfate
artificial substrate, Cys69 is the catalytic residue, and Lys302 and Lys123 act as residues anchoring the sulfate group to the active site, interaction with the enzyme active site, overview
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
-
135666, 135667, 135668, 135669, 135670, 135671, 135672, 135676, 135677, 135678, 135681, 135686, 135687, 135691, 135695, 649125
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
galactosyl ceramid
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
galactosyl ceramide
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
activity depends on ionic strength
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
no requirement for activators when acting on cerebroside sulfate
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
3-O-sulfogalactosyl ceramide
galactosyl ceramide
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
the enzyme catalyzes the first step in the degradation of the glycolipid cerebroside sulfate. Accumulation of cerebroside sulfate is responsible for metachromatic leukodystrophy, a severe autosomal recessive inherited disorder. Good correlation between the type of mutation, deficient arylsulfatase A activity and disease severity is recognized
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
the reaction requires saposin B, a non-enzymatic proteinaceous cofactor which presents sulfatide to the catalytic site of arylsulfatase A, desulfation is negligible when Sap B is substituted by Sap A, C, or D
-
-
?
p-nitrocatechol sulfate + H2O
p-nitrocatechol + sulfate
-
-
-
-
?
sulfogalactosylceramide + H2O
?
Cys69 is the catalytic residue, and Lys302 and Lys123 act as residues anchoring the sulfate group to the active site, interaction with the enzyme active site, overview
-
-
?
sulfogalactosylglycerolipid + H2O
?
Cys69 is the catalytic residue, and Lys302 and Lys123 act as residues anchoring the sulfate group to the active site, interaction with the enzyme active site, overview
-
-
?
additional information
?
-
-
metachromatic leukodystrophy is a lysosomal storage disorder caused by the deficiency of enzyme
-
-
?
additional information
?
-
substrate binding and docking study, using crystal structures, PDB IDs 1N2K and 1E2S, overview
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
cerebroside 3-sulphate + H2O
cerebroside + sulfate
the enzyme catalyzes the first step in the degradation of the glycolipid cerebroside sulfate
-
?
additional information
?
-
-
metachromatic leukodystrophy is a lysosomal storage disorder caused by the deficiency of enzyme
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
-
135666, 135667, 135668, 135669, 135670, 135671, 135672, 135676, 135677, 135678, 135681, 135686, 135687, 135691, 135695, 649125
-
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
galactosyl ceramid
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
-
galactosyl ceramide
-
?
cerebroside 3-sulfate + H2O
cerebroside + sulfate
the enzyme catalyzes the first step in the degradation of the glycolipid cerebroside sulfate. Accumulation of cerebroside sulfate is responsible for metachromatic leukodystrophy, a severe autosomal recessive inherited disorder. Good correlation between the type of mutation, deficient arylsulfatase A activity and disease severity is recognized
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
cerebroside sulfate
-
competitive to 1-O-acyl-2-O-alkyl3-O-(beta-D-galactopyranoside-3'-sulfate)glycerol
Na+
-
enzymatic activity is inhibited at NaCl concentrations exceeding 20 mM, at 50 and 200 mM the rate of sulfatide hydrolysis is diminished by about 50 and 75%, respectively
additional information
-
the lysosomal degradation of endocytosed enzymes strongly inhibited in CLN6 defective cells
-
additional information
-
estrogens do not significantly change ASA activity in epithelial, HCC1937, or MCF10A cells. Estrone 3-sulfate and estradiol sulfate do not significantly decline ASA in myoepithelial, MCF7, and T47D cells
-
additional information
-
secretion and stability of wtASA is decreased by the coexpression of defective P377L-pdASA
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
bis(monoacylglycero) phosphate
-
at a relative concentration of 30 mol% the lipid stimulates the reaction rate by a factor of 3.4
-
dolichol
-
at a relative concentration of 30 mol% the lipid stimulates the reaction rate by a factor of 2.7
phosphatidic acid
-
at a relative concentration of 30 mol% the lipid stimulates the reaction rate by a factor of 3.1
additional information
-
phosphatidylinositol and phosphatidylserine have no effect on the rate of sulfatide hydrolysis
-
Protein factor
-
activator protein stimulates lipid soluble substrates more than water soluble substrates
-
Protein factor
-
enzyme cleaves cerebroside sulfates only in the presence of either specific detergent, e.g. taurodeoxycholate or human activator protein
-
Protein factor
-
no requirement for activators when acting on cerebroside sulfate
-
Protein factor
-
degradation of sulfosphingolipids and sulfoglycerolipids with or without activator protein, critical micellar concentration important
-
taurodeoxycholate
-
enzyme cleaves cerebroside sulfates only in the presence of either specific detergent, e.g. taurodeoxycholate or human activator protein
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.2 - 0.5
1-O-Alkyl-2-O-acyl-3-O-(beta-D-galactopyranoside-3'-sulfate)-glycerol
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.001593
-
female with benign breast disease, comparison of activities at different ages
0.002233
-
female with breast cancer, comparison of activities at different ages
0.0148
-
arylsulfatase A co-expressed with formylglycine-generating enzyme, in liver
0.0272
-
arylsulfatase A co-expressed with formylglycine-generating enzyme, in serum
1593
-
woman with benign breast disease, comparison of activities at different ages
additional information
-
in HEK-293 cells, HeLa cells and COS-7 cells specific activity significantly increased when co-expressed with formylglycine-generating enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5
4.6
5.3
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3.75 - 4.75
-
desulfation activity is less than half-maximum at pH values below pH 3.75 and above pH 4.75
4 - 5.8
-
pH 4, pH 5.8: circa 20% of activity maximum, 10 mM sodium formate buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
135666, 135667, 135668, 135669, 135670, 135671, 135672, 135676, 135677, 135678, 135681, 135686, 135687, 135691
-
-
81 woman, including 28 healthy woman, 29 woman with benign breast disease and 24 patients with primary breast cancer
-
-
plasmid encoding human arylsulfatase A was expressed in arylsulfatase A-deficient mice, HEK-293 cells, HeLa cells and COS-7 cells
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
-
the polypeptide complexes, rather than the monomers, are subject to endoplasmic reticulum quality control and, within a heteromer, the misfolded subunit exerts a dominant negative effect on the wild-type subunit
-
the mean leukocyte enzyme activity in patients with benign breast disease is slightly higher than that (14.3%) that observed in the healthy subjects. In patients with primary breast cancer the enzyme activity is significantly higher than in healthy subjects
-
human fibroblasts grown in the presence of lead acetate exhibit a 65% decrease in enzyme protein
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
metabolism
-
arylsulfatase A catalyzes the first step in the intralysosomal degradation of the sphingolipid 3-O-sulfogalactosylceramide, briefly called sulfatide
physiological function
ARSA is involved in the catabolism of membrane sulfatides into galactosylceramide
additional information
-
heteromerization of wild-type and misfolded endoplasmic reticulum-degraded arylsulfatase A polypeptides affects the quality control of wild-type arylsulfatase A subunits. Within a heteromer, the misfolded subunit exerts a dominant negative effect on the wild-type subunit. Mechanism, overview
malfunction
-
metachromatic leukodystrophy, MLD, is a lysosomal storage disease caused by a deficiency of the lysosomal enzyme arylsulfatase A
malfunction
metachromatic leukodystrophy, MLD, is a rare inherited lysosomal storage disorder caused by the deficiency of arylsulfatase A, ARSA
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). ARSA_HUMAN
507
0
53588
Swiss-Prot
Secretory Pathway (Reliability: 1)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
49000
50000
55000
49000
-
aggregation to tetramer at higher concentrations around pH 5, 2 * 49000, 2 * 55000
50000
enzyme is synthesized as a preprotein with 507 amino acids, the mature protein has 489 amino acids with a calculated mass of 52000
55000
-
aggregation to tetramer at higher concentrations around pH 5, 2 * 49000, 2 * 55000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
octamer
oligomer
tetramer
-
aggregation to tetramer at higher concentrations around pH 5, 2 * 49000, 2 * 55000
additional information
octamer
-
ar pH 5.0-5.4, the enzyme exists in solution as a dimer or as an octamer
additional information
The dimer-octamer equilibrium is regulated by the pH and may be explained by a switch function of Glu424. Glu424 in the conformation suitable for the intramolecular hydrogen bonds to Gln460. Glu424 in the conformation suitable for the intermolecular hydrogen bonds to Phe398
additional information
-
mature lysosomal arylsulfatase A forms dimers at neutral pH, while in the early biosynthetic pathway, arylsulfatase A forms oligomers with more than two subunits. Within a heteromer, the misfolded subunit exerts a dominant negative effect on the wild-type subunit
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
phosphoprotein
-
compared to the degree of mannose-phosphorylation of the wild-type enzyme, the mutant enzyme K457R has 33%, mutant enzyme K457S has 50%, mutant enzyme K457G has 31%, mutant enzyme K433A has 95%, mutant enzyme K367A has 106% and mutant enzyme K393A has 123% of mannose-phosphorylation respectively
glycoprotein
-
the wild-type enzyme has three N-glycosylation sites. Only oligosaccharides at the first, Asn158, and the third, Asn350, glycosylation site are phosphorylated, whereas the second, Asn184 is not
glycoprotein
-
three N-glycosylation sites and three N-linked oligosaccharide side chains at asparagine residues 158, 184, and 350, the recombinantly expressed enzyme shows a high variability of the high-mannose-type N-glycans, which prevail at all glycosylation sites, depending on the culture conditions and the cell line expressing the enzyme, overview. The composition of the glycans is largely determined by substantial trimming in the medium, the susceptibility for trimming is different for the glycans at the three N-glycosylation sites, which of the glycans is most susceptible to trimming also depends on production conditions. Structure and ligand binding analysis, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of mutant C69A a nd C69S in complex with p-nitrocatechol sulfate
crystal structure of mutant C69A in complex with p-nitrocatechol sulfate, Protein Data Bank: 1E1Z and 1E2S, and crystal structure of mutant C69S in complex with p-nitrocatechol sulfate, Protein Data Bank: 1E33 and 1E3C
crystallisation of enzyme at pH 5.4 in a new monoclinic crystal form, at pH 6.5-6.7, tetragonal crystals are obtained.
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A212P
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
C168stop
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
C300F
-
the sequence alteration is found in a patient with metachromatic leukodystrophy, the mutant strongly interferes with the octamerization process of enzyme but not with its dimerization capacity
C500F
C69A
The inactive mutant in complex with p-nitrocatechol sulfate mimics a reaction intermediate during sulfate ester hydrolysis by the active enzyme, without the covalent bond to the key side-chain C-alpha-formylglycine
C69S
D407fs
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
E253K/T391S
mutations contribute to sum of the enzyme activity reduction ascribed to each mutation
E307K
naturally occuring ARSA polymorphism, causes a mild peripheral neuropathy phenotype
E382Q
H138D
naturally occuring ARSA polymorphism, causes a mild peripheral neuropathy phenotype
H231Q
mutation identified in three patients belonging to a consanguineous family with late-infantile metachromatic leukodystrophy disorder MLD. The mutation leads to changes in the pre-mRNA secondary structure and in the ArsA protein structure
K302A
K367A
K393A
K393A/K395G
K393A/K395H
K433A
K457G
K457R
K457S
K463Q
K463R
L52P
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
N158Q/N350Q
N350S
P425T
-
the sequence alteration is found in a patient with metachromatic leukodystrophy, the mutant displays a modest reduction of oligomerization process but not with its dimerization capacity
P426L/N350S/96A>G
mutations contribute to sum of the enzyme activity reduction ascribed to each mutation
R288H
R288H/R496H/N350S
-
about 38% reduction of enzyme activity in comparison to wild-type enzyme, no additive effect of the various amino acid substitutions is found in vitro
R496H
S406G
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
T304M
naturally occuring ARSA polymorphism, causes a severe peripheral neuropathy phenotype
additional information
C500F
-
about 8% reduction of enzyme activity in comparison to wild-type enzyme. Mutation found in a patient with metachromatic leukodystrophy
C69S
the mutant is able to bind covalently to the substrate and hydrolyse it, but is unable to release the resulting sulfate
C69S
present at significantly higher levels in both conditioned media and within the cell when compared with wild type
E382Q
-
complete loss of enzyme activity in comparison to wild-type enzyme in vitro
E382Q
-
complete loss of enzyme activity in comparison to wild-type enzyme in vitro. Mutation found in a patient with metachromatic leukodystrophy
K302A
-
34% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium
K367A
-
60% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium. 106% of the mannose-phosphorylation of the wild-type enzyme
K393A
-
39% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium. 123% of the mannose-phosphorylation of the wild-type enzyme
K393A/K395G
-
35% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium
K393A/K395H
-
30% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium
K433A
-
102% of the activity of wild-type enzyme when expressed in BHK cells. Mutation has no effect on distribution of the enzyme activity between cell and medium. 95% of the mannose-phosphorylation of the wild-type enzyme
K457G
-
7.5% of the activity of wild-type enzyme when expressed in BHK cells. Mutation leads to an increase of enzyme activity in the medium to about 60% of total activity compared to 35% of the wild-type enzyme. 31% of the mannose-phosphorylation of the wild-type enzyme. Mutation affects phosphorylation of oligosaccharide Asn350 to a greater extent than that of oligosaccharide Asn158
K457R
-
45% of the activity of wild-type enzyme when expressed in BHK cells. Mutation leads to an increase of enzyme activity in the medium to about 60% of total activity compared to 35% of the wild-type enzyme. 33% of the mannose-phosphorylation of the wild-type enzyme. Mutation affects phosphorylation of oligosaccharide Asn350 to a greater extent than that of oligosaccharide Asn158
K457S
-
25% of the activity of wild-type enzyme when expressed in BHK cells. 50% of the mannose-phosphorylation of the wild-type enzyme. Mutation leads to an increase of enzyme activity in the medium to about 65% of total activity compared to 35% of the wild-type enzyme. 50% of the mannose-phosphorylation of the wild-type enzyme. Mutation affects phosphorylation of oligosaccharide Asn350 to a greater extent than that of oligosaccharide Asn158
K463Q
-
14% of the activity of wild-type enzyme when expressed in BHK cells. Mutation leads to an decrease of enzyme activity in the medium to about 25% of total activity compared to 35% of the wild-type enzyme
K463R
-
39% of the activity of wild-type enzyme when expressed in BHK cells. 31% of the mannose-phosphorylation of the wild-type enzyme. Mutation leads to an decrease of enzyme activity in the medium to about 25% of total activity compared to 35% of the wild-type enzyme
N158Q/N350Q
-
43% of the activity of wild-type enzyme when expressed in BHK cells. Mutation leads to an increase of enzyme activity in the medium to about 25% of total activity compared to 75% of the wild-type enzyme
N350S
-
about 15% reduction of enzyme activity in comparison to wild-type enzyme in vitro
N350S
-
about 15% reduction of enzyme activity in comparison to wild-type enzyme in vitro, mutation found in a patient with metachromatic leukodystrophy
R288H
-
about 40% reduction of enzyme activity in comparison to wild-type enzyme in vitro
R288H
-
about 40% reduction of enzyme activity in comparison to wild-type enzyme in vitro, mutation found in a patient with metachromatic leukodystrophy
R496H
-
about 15% reduction of enzyme activity in comparison to wild-type enzyme in vitro
R496H
-
about 15% reduction of enzyme activity in comparison to wild-type enzyme in vitro, mutation found in a patient with metachromatic leukodystrophy
additional information
-
R288H, N350S and R496H are sequence alterations found in a patient with metachromatic leukodystrophy. W124ter mutation, which leads to a truncated enzyme and the mutations E382Q and C500F are found in an other patient.
additional information
-
only substitution of Lys457 causes a reduction of phosphorylation to 33% and increases secretion of the mutant enzyme
additional information
-
R288H, N350S and R496H are sequence alterations found in a patient with metachromatic leukodystrophy. W124ter mutation, which leads to a truncated enzyme and the mutations E382Q and C500F are found in an other patient
additional information
study of disease-causing mutations, severe phenotype of this patient depends not only on the two disease-causing mutations, but also to some extent on the pseudodeficiency mutations
additional information
the frameshift mutations g.445_446dupG and g.2590_2591dupC are associated with metachromatic leukodystrophy and lead 0.3% or 10% of wild type ARSA activity, respectively
additional information
-
the frameshift mutations g.445_446dupG and g.2590_2591dupC are associated with metachromatic leukodystrophy and lead 0.3% or 10% of wild type ARSA activity, respectively
additional information
-
secretion and half-life of wild-type enzyme are reduced upon co-expression of defective P377L-pdASA
additional information
characterization of pathogenic variants c.98T > C, c.195delC, c.229G > C, c.545C > G, c.674A > G, c.852T > A, and c.1274A > G, which were found when sequencing a cohort of 31 German metachromatic leukodystrophy families. Upon expression in immortalized, human multipotent mesenchymal stromal cells prepared from a patient deficient in ARSA activity, the seven mutants show ARSA activity of less than 10% when compared with wild type
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, addition of dithiothreitol, glutathione, or EDTA reduce storage stability
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, pH 7.5, protein: 0.002 mg/ml, bovine serum albumin, insulin or ribonuclease, 15 days, no loss of activity
-
4°C, pH 7.5, protein: 0.002 mg/ml, bovine serum albumin, insulin or ribonuclease, 8 months, 30% loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
of the recombinant protein
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
coexpression and secretion of misfolded, enzymatically inactive enzyme with the wild-type enzyme, whose activity is diminished upon coexpression with misfolded ASA polypeptides, in BHK cells
-
expression in arylsulfatase A-deficient mice, HEK-293 cells, HeLa cells and COS-7 cells
-
expression in BHK cells
HSPC cells transduced with increasing doses of lentiviral vectors expressing ARSA under the control of the human PGK promoter (PGK.ARSA.LV). Transgenic mice generated with PGK.ARSA.LV overexpressing the ARSA enzyme
-
human arylsulfatase A tagged with hemagglutinin or GFP cloned in a lentiviral vector, expressed in HeLa cells and mice
-
plasmids containing the wild-type and mutant cDNAs are transiently transfected into BHK cells
-
recombinant expression in CHO, BHK and HT-1080 cells. High variability of the high-mannose-type N-glycans, which prevail at all glycosylation sites, depending on the culture conditions and the cell line expressing the enzyme, overview
-
the ARSA gene is a small gene located on chromosome 22q13 that spans 8 exons and encodes a 507-aminoacid peptide, it is transcribed into three mRNA species, a major one of 2.1 kb and two less abundant species of 3.7 and 4.8 kb. DNA and amino acid sequence determination and analysis of natural mutants. Recombinant expression in HeLa cells using lentiviral vectors, containing mutated ARSA alleles, the vectors stably integrate into the host genome. expression of mutated alleles in murine ARSA-/- fibroblasts
transient expression experiments on COS7 cells transfected with enzyme cDNAs carrying the mutations separately and in the combination found in the patient's alleles
transient expression experiments on COS7 cells transfected with enzyme cDNAs carrying the mutations separately and in the combination found in the patients alleles
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
-
deficiency in metachromatic leukodystrophy, a sphingolipid storage disorder
medicine
-
assay of activity in leukocytes as a non-invasive diagnostic tool in patients with benign and malignant breast disease
medicine
-
metachromatic leukodystrophy is a lysosomal storage disorder caused by the deficiency of enzyme
medicine
-
the enzyme is implicated in most cases of metachromatic leukodystrophy
medicine
-
therapeutic levels of ARSA overexpression can be safetly achieved. ARSA-transduced cells efficiently repopulate the hematopoietic organs of RAG2-/- gamma-chain-/- mice. ARSA overexpression does not impair clonogenic capacity and multilineage differentiation of human HSPC cells, activation of other sulfatases and is thus unlikely to trigger metabolic imbalances. Moreover, ARSA overexpression does not impair immune functions, behavior and learning abilities
medicine
-
tyrosine sulfation facilitates secretion of ASA, which may have pathophysiological consequences
medicine
metachromatic leukodystrophy is caused by deficient activity of arylsulfatase A
medicine
-
enzyme replacement therapy is a therapeutic option for metachromatic leukodystrophy, caused by enzyme-deficiency, and other lysosomal disorders. This therapy depends on N-linked oligosaccharide-mediated delivery of intravenously injected recombinant enzyme to the lysosomes of patient cells
medicine
characterization of pathogenic variants C490R, P428L, N284K, P428L, H425R, Y225C and P428L, which were found when sequencing a cohort of 31 German metachromatic leukodystrophy families. Upon expression in immortalized, human multipotent mesenchymal stromal cells prepared from a patient deficient in ARSA activity, the seven mutants show ARSA activity of less than 10% when compared with wild type
medicine
mutation H231Q occurs in three patients belonging to a consanguineous family with late-infantile metachromatic leukodystrophy disorder MLD. The mutation leads to changes in the pre-mRNA secondary structure and in the ArsA protein structure
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Sarafian, T.A.; Fluharty, A.L.; Kihara, H.; Helfand, G.; Edmond, J.
Large scale purification of pyrogen-free human arylsulfatase A
J. Appl. Biochem.
4
126-132
1982
Homo sapiens
-
Fluharty, A.L.; Stevens, R.L.; Miller, R.T.; Kihara, H.
Bile salt activation of cerebroside sulphate sulphohydrolase
Biochem. J.
189
45-49
1980
Homo sapiens
Fischer, G.; Jatzkewitz, H.
The activator of cerebroside-sulphatase. A model of the activation
Biochim. Biophys. Acta
528
69-76
1978
Homo sapiens
Stevens, R.L.; Fluharty, A.L.; Killgrove, A.R.; Kihara, H.
Microheterogeneity of arylsulfatase A from human tissues
Biochim. Biophys. Acta
445
661-671
1976
Homo sapiens
Fischer, G.; Jatzkewitz, H.
The activator of cerebroside sulphatase. Purification from human liver and identification as a protein
Hoppe-Seyler's Z. Physiol. Chem.
356
605-613
1975
Homo sapiens
Mraz, W.; Fischer, G.; Jatzkewitz, H.
The activator of human cerebroside sulphatase. Activating effect on the acidic forms of the sulphatases from invertebrates
Hoppe-Seyler's Z. Physiol. Chem.
357
201-206
1976
Homo sapiens, Maja squinado, Marthasterias glacialis, Microcosmus sulcatus, Patella vulgata, Tethya aurantium
Mraz, W.; Fischer, G.; Jatzkewitz, H.
The activator of cerebroside sulphatase. Lysosomal localization
Hoppe-Seyler's Z. Physiol. Chem.
357
1181-1191
1976
Homo sapiens, Rattus norvegicus
Stinshoff, K.; Jatzkewitz, H.
Comparison of the cerebroside sulphatase and the arylsulphatase activity of human sulphatase A in the absence of activators
Biochim. Biophys. Acta
377
126-138
1975
Homo sapiens
Fluharty, A.L.; Stevens, R.L.; Miller, R.T.; Kihara, H.
Sulfoglycerogalactolipid from rat testis: a substrate for pure human arylsulfatase A
Biochem. Biophys. Res. Commun.
61
348-354
1974
Homo sapiens
Porter, M.T.; Fluharty, A.L.; De la Flor, S.; Kihara, H.
Cerebroside sulfatase determination in cultured human fibroblasts
Biochim. Biophys. Acta
258
769-778
1972
Homo sapiens
Fluharty, A.L.; Edmond, J.
Arylsulfatases A and B from human liver
Methods Enzymol.
50
537-547
1978
Homo sapiens
Stevens, R.L.; Fluharty, A.L.; Skokut, M.H.; Kihara, H.
Purification and properties of arylsulfatase. A from human urine
J. Biol. Chem.
250
2495-2501
1975
Homo sapiens
Fischer, G.; Reiter, S.; Jatzkewitz, H.
Enzymic hydrolysis of sulphosphingolipids and sulphoglycerolipids by sulphatase A in the presence and absence of activator protein
Hoppe-Seyler's Z. Physiol. Chem.
359
863-866
1978
Homo sapiens
Park, D.S.; Poretz, R.D.; Ricketts, M.H.; Manowitz, P.
Arylsulfatase A: relationship of genotype to variant electrophoretic properties
Biochem. Genet.
34
149-161
1996
Homo sapiens
Lukatela, G.; Krauss, N.; Theis, K.; Selmer, T.; Gieselmann, V.; von Figura, K.; Saenger, W.
Crystal structure of human arylsulfatase A: the aldehyde function and the metal ion at the active site suggest a novel mechanism for sulfate ester hydrolysis
Biochemistry
37
3654-3664
1998
Homo sapiens (P15289)
Sommerlade, H.J.; Selmer, T.; Ingendoh, A.; Gieselmann, V.; von Figura, K.; Neifer, K.; Schmidt, B.
Glycosylation and phosphorylation of arylsulfatase A
J. Biol. Chem.
269
20977-20981
1994
Homo sapiens
Roy, A.B.
Sulphatases, lysosomes and disease
Aust. J. Exp. Biol. Med. Sci.
54
111-135
1976
Bos taurus, Oryctolagus cuniculus, Homo sapiens
Franco, B.; Meroni, G.; Parenti, G.; Levilliers, J.; Bernard, L.; Gebbia, M.; Cox, L.; Maroteaux, P.; Sheffield, L.; Rappold, G.A.; et al.
A cluster of sulfatase genes on Xp22.3: mutations in chondrodysplasia punctata (CDPX) and implications for warfarin embryopathy
Cell
81
15-25
1995
Homo sapiens
Lewinski, K.; Chruszcz, M.; Ksiazek, D.; Laidler, P.
Crystallization and preliminary crystallographic analysis of a new crystal form of arylsulfatase A isolated from human placenta
Acta Crystallogr. Sect. D
56
650-652
2000
Homo sapiens
-
Marcao, A.; Simonis, H.; Schestag, F.; Sa Miranda, M.C.; Gieselmann, V.
Biochemical characterization of two (C300F, P425T) arylsulfatase a missense mutations
Am. J. Med. Genet.
116A
238-242
2003
Homo sapiens
-
Vagedes, P.; Saenger, W.; Knapp, E.W.
Driving forces of protein association: the dimer-octamer equilibrium in arylsulfatase A
Biophys. J.
83
3066-3078
2002
Homo sapiens (P15289)
Turkmen, S.; Oner, P.; Cinar, F.; Kocak, H.; Guvenen, G.; Altun, H.; Eryavuz, Y.
Evaluation of leukocyte arylsulfatase-A activity in patients with breast cancer and benign breast disease
Cancer Lett.
166
95-101
2001
Homo sapiens
Regis, S.; Corsolini, F.; Stroppiano, M.; Cusano, R.; Filocamo, M.
Contribution of arylsulfatase A mutations located on the same allele to enzyme activity reduction and metachromatic leukodystrophy severity
Hum. Genet.
110
351-355
2002
Homo sapiens (P15289)
Yaghootfam, A.; Schestag, F.; Dierks, T.; Gieselmann, V.
Recognition of arylsulfatase A and B by the UDP-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-phosphotransferase
J. Biol. Chem.
278
32653-32661
2003
Homo sapiens
Heine, C.; Koch, B.; Storch, S.; Kohlschutter, A.; Palmer, D.N.; Braulke, T.
Defective endoplasmic reticulum-resident membrane protein CLN6 affects lysosomal degradation of endocytosed arylsulfatase A
J. Biol. Chem.
279
22347-22352
2004
Homo sapiens
von Bulow, R.; Schmidt, B.; Dierks, T.; von Figura, K.; Uson, I.
Crystal structure of an enzyme-substrate complex provides insight into the interaction between human arylsulfatase A and its substrates during catalysis
J. Mol. Biol.
305
269-277
2001
Homo sapiens (P15289)
Yaghootfam, A.; Baumann, N.; Schwarz, A.; Gieselmann, V.
Three novel mutant arylsulfatase A alleles causing metachromatic leukodystrophy
Neurochem. Res.
29
933-942
2004
Homo sapiens
Chen, X.G.; Poretz, R.D.
Lead causes human fibroblasts to mis-sort arylsulfatase A
Toxicology
163
107-114
2001
Homo sapiens
Christianson, T.M.; Starr, C.M.; Zankel, T.C.
Overexpression of inactive arylsulfatase mutants and in vitro activation by light-dependent oxidation with vanadate
Biochem. J.
382
581-587
2004
Homo sapiens (P15289)
Bognar, S.K.; Foretic, B.; Furac, I.; Vukelic, Z.; Grubesic, Z.
Kinetics and activity of arylsulfatase A in leukocytes derived from patients with cerebral palsy
Acta Pharm.
56
95-104
2006
Homo sapiens
Takakusaki, Y.; Hisayasu, S.; Hirai, Y.; Shimada, T.
Coexpression of formylglycine-generating enzyme is essential for synthesis and secretion of functional arylsulfatase A in a mouse model of metachromatic leukodystrophy
Hum. Gene Ther.
16
929-936
2005
Homo sapiens
Luca, T.; Givogri, M.I.; Perani, L.; Galbiati, F.; Follenzi, A.; Naldini, L.; Bongarzone, E.R.
Axons mediate the distribution of arylsulfatase A within the mouse hippocampus upon gene delivery
Mol. Ther.
12
669-679
2005
Homo sapiens
Oner-Iyidogan, Y.; Oner, P.; Kocak, H.; Lama, A.; Gurdol, F.; Bekpinar, S.; Unur, N.; Ozbek-Kir, Z.
Evaluation of leukocyte arylsulphatase a, serum interleukin-6 and urinary heparan sulphate following tamoxifen therapy in breast cancer
Pharmacol. Res.
52
340-345
2005
Homo sapiens
Bhattacharyya, S.; Tobacman, J.K.
Steroid sulfatase, arylsulfatases A and B, galactose-6-sulfatase, and iduronate sulfatase in mammary cells and effects of sulfated and non-sulfated estrogens on sulfatase activity
J. Steroid Biochem. Mol. Biol.
103
20-34
2007
Homo sapiens
Kasinathan, C.; Jean, S.; Manowitz, P.
Tyrosine sulfation of arylsulfatase A and its peptide
Protein Pept. Lett.
13
357-361
2006
Homo sapiens
Capotondo, A.; Cesani, M.; Pepe, S.; Fasano, S.; Gregori, S.; Tononi, L.; Venneri, M.A.; Brambilla, R.; Quattrini, A.; Ballabio, A.; Cosma, M.P.; Naldini, L.; Biffi, A.
Safety of arylsulfatase A overexpression for gene therapy of metachromatic leukodystrophy
Hum. Gene Ther.
18
821-836
2007
Homo sapiens
Lugowska, A.; Wlodarski, P.; Ploski, R.; Mierzewska, H.; Dudzinska, M.; Matheisel, A.; Swietochowska, H.; Tylki-Szymanska, A.
Molecular and clinical consequences of novel mutations in the arylsulfatase A gene
Clin. Genet.
75
57-64
2009
Homo sapiens (P15289), Homo sapiens
Matzner, U.; Breiden, B.; Schwarzmann, G.; Yaghootfam, A.; Fluharty, A.L.; Hasilik, A.; Sandhoff, K.; Gieselmann, V.
Saposin B-dependent reconstitution of arylsulfatase a activity in vitro and in cell culture models of metachromatic leukodystrophy
J. Biol. Chem.
284
9372-9381
2009
Homo sapiens
Schroeder, S.; Matthes, F.; Hyden, P.; Andersson, C.; Fogh, J.; Mueller-Loennies, S.; Braulke, T.; Gieselmann, V.; Matzner, U.
Site-specific analysis of N-linked oligosaccharides of recombinant lysosomal arylsulfatase A produced in different cell lines
Glycobiology
20
248-259
2010
Homo sapiens
Schenk, M.; Koppisetty, C.A.; Santos, D.C.; Carmona, E.; Bhatia, S.; Nyholm, P.G.; Tanphaichitr, N.
Interaction of arylsulfatase-A (ASA) with its natural sulfoglycolipid substrates: a computational and site-directed mutagenesis study
Glycoconj. J.
26
1029-1045
2009
Homo sapiens (P15289), Sus scrofa (Q8WNR3)
Cesani, M.; Capotondo, A.; Plati, T.; Sergi, L.S.; Fumagalli, F.; Roncarolo, M.G.; Naldini, L.; Comi, G.; Sessa, M.; Biffi, A.
Characterization of new arylsulfatase A gene mutations reinforces genotype-phenotype correlation in metachromatic leukodystrophy
Hum. Mutat.
30
E936-E945
2009
Homo sapiens (P15289)
Poeppel, P.; Abouzied, M.M.; Voelker, C.; Gieselmann, V.
Misfolded endoplasmic reticulum retained subunits cause degradation of wild-type subunits of arylsulfatase A heteromers
FEBS J.
277
3404-3414
2010
Homo sapiens
Boehringer, J.; Santer, R.; Schumacher, N.; Gieseke, F.; Cornils, K.; Pechan, M.; Kustermann-Kuhn, B.; Handgretinger, R.; Schoels, L.; Harzer, K.; Kraegeloh-Mann, I.; Mueller, I.
Enzymatic characterization of novel arylsulfatase A variants using human arylsulfatase A-deficient immortalized mesenchymal stromal cells
Hum. Mutat.
38
1511-1520
2017
Homo sapiens (P15289)
Issa, A.B.; Feki, F.K.; Jdila, M.B.; Khabou, B.; Rhouma, B.B.; Ammar-Keskes, L.; Triki, C.; Fakhfakh, F.
Clinical, molecular, and computational analysis showed a novel homozygous mutation among the substrate-binding site of ARSA protein in consanguineous family with late-infantile MLD
J. Mol. Neurosci.
66
17-25
2018
Homo sapiens (P15289)
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