Information on EC 3.2.1.49 - alpha-N-acetylgalactosaminidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.2.1.49
-
RECOMMENDED NAME
GeneOntology No.
alpha-N-acetylgalactosaminidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Cleavage of non-reducing alpha-(1->3)-N-acetylgalactosamine residues from human blood group A and AB mucin glycoproteins, Forssman hapten and blood group A lacto series glycolipids
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycosphingolipid biosynthesis - globo series
-
-
SYSTEMATIC NAME
IUBMB Comments
alpha-N-acetyl-D-galactosaminide N-acetylgalactosaminohydrolase
The human lysosomal enzyme is involved in the degradation of blood type A epitope.
CAS REGISTRY NUMBER
COMMENTARY hide
9075-63-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
squid, two isozymes alpha-GalNAcase I and alpha-GalNAcase II
-
-
Manually annotated by BRENDA team
isolated from brown algae Chorda filum
-
-
Manually annotated by BRENDA team
isolated from brown algae Chorda filum
-
-
Manually annotated by BRENDA team
KMM 426
-
-
Manually annotated by BRENDA team
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Arenibacter sp.
isolated from green algae Acrosiphonia sonderi
-
-
Manually annotated by BRENDA team
Arenibacter sp. KMM 6041
isolated from green algae Acrosiphonia sonderi
-
-
Manually annotated by BRENDA team
strain CCIM K2, best producer of alpha-N-acetylgalactosaminase
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Cellulophaga sp.
isolated from brown algae Acrosiphonia sonderi; isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
Cellulophaga sp. KMM 6483
isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
Cellulophaga sp. KMM 6488
isolated from brown algae Acrosiphonia sonderi
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Eremoplastron bovis
-
-
-
Manually annotated by BRENDA team
Formosa sp.
isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
Formosa sp. KMM 6408
isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
i.e. HIV-1, constitutive enzyme
-
-
Manually annotated by BRENDA team
skipjack tuna
-
-
Manually annotated by BRENDA team
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Maribacter sp.
isolated from green algae Acrosiphonia sonderi; isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 3952
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 6484
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 6485
isolated from green algae Acrosiphonia sonderi
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 6486
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 6487
isolated from green algae Acrosiphonia sonderi
-
-
Manually annotated by BRENDA team
Maribacter sp. KMM 6489
isolated from green algae Ulva fenestrata
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
shrimp
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
squid
-
-
Manually annotated by BRENDA team
isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
isolated from brown algae Saccharina japonica
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
additional information
-
comparison of the binding ability of alpha-N-acetylgalactosaminidase with red blood cells in different reaction buffers, such as normal saline, phosphate-buffered saline, a disodium hydrogen phosphate-based buffer, and 5% commercial glucose solution. Glucose buffer is suitable for blood group conversion with alpha-N acetylgalactosaminidase
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-nitrophenyl 2-acetyl-2-deoxy-alpha-D-galactosamine + H2O
2-nitrophenol + 2-acetyl-2-deoxy-alpha-D-galactosamine
show the reaction diagram
2-nitrophenyl-alpha-D-galactoside + H2O
2-nitrophenol + alpha-D-galactose
show the reaction diagram
-
-
-
?
2-nitrophenyl-alpha-N-acetylgalactosaminide + H2O
2-nitrophenol + alpha-N-acetylgalactosamine
show the reaction diagram
4-methylumbelliferyl-alpha-D-galactopyranoside + H2O
4-methylumbelliferone + alpha-D-galactopyranose
show the reaction diagram
4-methylumbelliferyl-alpha-D-galactosaminide + H2O
4-methylumbelliferone + alpha-D-galactosamine
show the reaction diagram
-
-
-
-
?
4-nitrophenyl alpha-D-N-acetylgalactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
show the reaction diagram
4-nitrophenyl N-acetyl-alpha-D-galactosamine + H2O
4-nitrophenol + N-acetyl-alpha-D-galactosamine
show the reaction diagram
4-nitrophenyl N-acetyl-alpha-D-galactosamine + H2O
4-nitrophenol + N-acetyl-alpha-galactosamine
show the reaction diagram
-
-
-
-
?
4-nitrophenyl N-acetyl-beta-D-galactosamine + H2O
4-nitrophenol + N-acetyl-beta-galactosamine
show the reaction diagram
-
-
-
-
?
4-nitrophenyl-N-acetyl-alpha-D-galactosamide + H2O
4-nitrophenol + N-acetyl-alpha-D-galactosamine
show the reaction diagram
-
-
-
-
?
blood group A glycoconjugate + H2O
?
show the reaction diagram
Forssman hapten + H2O
?
show the reaction diagram
GalNAc-alpha-1,3-GalNAc-beta-1,3-Gal-alpha-1,4-Gal-beta-1,4-Glc-beta-1,1-ceramide + H2O
alpha-N-acetylgalactosamine + GalNAc-beta-1,3-Gal-alpha-1,4-Gal-beta-1,4-Glc-beta-1,1-ceramide
show the reaction diagram
GalNAc-alpha1-O-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
GalNAc-alpha1-O-Thr + H2O
?
show the reaction diagram
-
-
-
-
?
globotriaosylceramide + H2O
?
show the reaction diagram
wild-type and engineered mutant enzymes
-
-
?
o-nitrophenyl-alpha-N-acetylgalactosaminide + H2O
o-nitrophenol + alpha-N-acetylgalactosamine
show the reaction diagram
o-nitrophenyl-N-acetyl-alpha-D-galactosaminide + H2O
o-nitrophenol + N-acetyl-alpha-D-galactosamine
show the reaction diagram
ovine submaxillary asialoglycoprotein + H2O
?
show the reaction diagram
-
-
-
-
?
p-nitrophenyl 2-acetamide-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside + H2O
p-nitrophenol + 2-acetamide-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside
show the reaction diagram
-
-
-
-
?
p-nitrophenyl alpha-D-galactopyranoside + H2O
p-nitrophenol + alpha-D-galactopyranose
show the reaction diagram
-
-
-
-
?
p-nitrophenyl N-acetyl-alpha-D-galactosaminide + H2O
p-nitrophenol + N-acetyl-alpha-D-galactosamine
show the reaction diagram
-
-
-
-
?
p-nitrophenyl N-acetyl-alpha-D-galactosaminide + H2O
p-nitrophenol + N-acetyl-D-galactosamine
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-2-deoxy-alpha-D-galactopyranoside + H2O
p-nitrophenol + 2-deoxy-alpha-D-galactopyranose
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-alpha-N-acetyl-D-galactoaminide + H2O
p-nitrophenol + alpha-N-acetyl-D-galactosamine
show the reaction diagram
-
the Hep-G2 cell lysate hydrolyzes the exo-type substrate
-
-
?
p-nitrophenyl-alpha-N-acetyl-D-galactosaminide + H2O
p-nitrophenol + alpha-N-acetyl-D-galactosamine
show the reaction diagram
p-nitrophenyl-alpha-N-acetylgalactosaminide + H2O
p-nitrophenol + alpha-N-acetylgalactosamine
show the reaction diagram
p-nitrophenyl-N-acetyl-alpha-D-galactosaminide + H2O
p-nitrophenol + N-acetyl-alpha-D-galactosamine
show the reaction diagram
phenyl-alpha-N-acetylglucosaminide + H2O
N-acetylgalactosamine + phenol
show the reaction diagram
-
-
-
?
serum vitamin D3-binding protein + H2O
?
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
blood group A glycoconjugate + H2O
?
show the reaction diagram
-
alpha-N-acetylgalactosaminidase blood group A2 degrading activity, substrate from human erythrocytes and plasma
-
-
?
serum vitamin D3-binding protein + H2O
?
show the reaction diagram
-
i.e. Gc protein, the human substrate is the precursor of the principal macrophage activating factor, MAF, the deglycosylated prrecursor cannot be converted, which leads to immunosuppression
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
binds 1 NAD+ per subunit. The NAD+ cannot dissociate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
slight activation of both isozymes
Cu2+
-
slight activation of both isozymes
K+
-
slight activation of both isozymes
Na+
-
slight activation of both isozymes
Zn2+
-
slight activation of both isozymes
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dianisidine
-
2 mM, 70% of original activity
Acetylimidazole
-
5 mM, 30% of original activity
carbodiimide
-
1.8 mM, complete loss of activity
D-galactose
D-glucose
-
-
Diethylpyrocarbonate
-
2 mM, 60% of original activity
Fe2+
-
-
Mg2+
-
-
Mn2+
-
-
N-acetyl-D-galactosamine
N-acetyl-D-glucosamine
-
-
N-acetylgalactosamine
N-bromosuccinimide
-
1.8 mM, complete loss of activity
N-ethylmaleimide
-
1.8 mM, 60% of original activity
p-chloromercuribenzoate
-
1.8 mM, complete loss of activity
Pb2+
-
10 mM, 30% residual activity
Zn2+
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Trypsin
-
the enzyme needs to be proteolytically activated, activation of the enzyme by trypsin
-
additional information
-
the enzyme is inducible in patients peripheral blood mononuclear cells by HIV-1 provirus-inducing agent, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.56 - 0.73
2-nitrophenyl-alpha-N-acetylgalactosaminide
6.8
4-methylumbelliferyl-alpha-D-galactopyranoside
-
-
0.7 - 0.89
4-nitrophenyl alpha-D-N-acetylgalactosaminide
0.036 - 0.59
Forssman hapten
1.04 - 3.97
GalNAc-alpha-1,3-GalNAc-beta-1,3-Gal-alpha-1,4-Gal-beta-1,4-Glc-beta-1,1-ceramide
7
N-acetyl-alpha-D-galactosaminide
-
-
8.8
o-nitrophenyl-alpha-D-fucopyranoside
-
-
1.3 - 2.4
o-nitrophenyl-alpha-N-acetylgalactosaminide
1.58 - 1.62
o-nitrophenyl-N-acetyl-alpha-D-galactosaminide
0.05
ovine submaxillary asialoglycoprotein
-
-
-
1.27 - 4.83
p-nitrophenyl N-acetyl-alpha-D-galactosaminide
14.7
p-nitrophenyl-2-deoxy-alpha-D-galactopyranoside
-
-
0.38
p-nitrophenyl-alpha-N-acetyl-D-galactosaminide
-
pH 7.2, 36C
0.23 - 6.6
p-nitrophenyl-alpha-N-acetylgalactosaminide
1.1 - 1.35
p-nitrophenyl-N-acetyl-alpha-D-galactosaminide
50
phenyl-alpha-N-acetylglucosaminide
-
-
1.27 - 4.83
serum vitamin D3-binding protein
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.3 - 17.1
4-nitrophenyl alpha-D-N-acetylgalactosaminide
19.7
4-nitrophenyl N-acetyl-alpha-D-galactosamine
Elizabethkingia meningoseptica
-
recombinant enzyme, pH 7.5, 37C
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1250
4-nitrophenyl N-acetyl-alpha-D-galactosamine
Elizabethkingia meningoseptica
-
recombinant enzyme, pH 7.5, 37C
202487
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0002
-
cell extract of gingival keratinocyte
0.0003
-
cell extract of gingival fibroblast
0.0004
-
cell extract of embryonic lung fibroblast
0.0012
-
pH 5.0, Chang liver cell
0.0021
-
pH 5.0, Hep-G2 cell
0.0026
-
pH 5.0, HCT116 cell
0.005
-
cell extract of gingival carcinoma
0.012
-
cell extract of salivary gland carcinoma
0.63
Maribacter sp.
-
crude enzyme, strain KMM 6486, pH 7.3, 20C
0.64
-
crude enzyme, pH 7.3, 20C
0.65
Maribacter sp.
-
crude enzyme, strain KMM 6484, pH 7.3, 20C
0.79
Maribacter sp.
-
crude enzyme, strain KMM 6489, pH 7.3, 20C
0.83
-
crude enzyme, strain KMM 3961, pH 7.3, 20C
0.84
Arenibacter sp.
-
crude enzyme, pH 7.3, 20C
0.91
Maribacter sp.
-
crude enzyme, strain KMM 6485, pH 7.3, 20C
0.92
Formosa sp.
-
crude enzyme, pH 7.3, 20C
1.38
Maribacter sp.
-
crude enzyme, strain KMM 3952, pH 7.3, 20C
1.7
-
crude enzyme, strain KMM 3528, pH 7.3, 20C
1.8
Cellulophaga sp.
-
crude enzyme, strain KMM 6483, pH 7.3, 20C
1.83
-
crude enzyme, strain KMM 3980, pH 7.3, 20C
3.1
substrate 2-nitrophenyl-alpha-D-galactoside, pH not specified in the publication, temperature not specified in the publication
4.5
-
purified placental enzyme
5.1
-
crude enzyme, strain KMM 3523, pH 7.3, 20C
5.5
Cellulophaga sp.
-
crude enzyme, strain KMM 6488, pH 7.3, 20C
8.33
-
purified recombinant enzyme mutant
19.56
-
purified isozyme alpha-GalNAcase I
24.2
-
purified isozyme alpha-GalNAcase II
42.3
pH 3.5, 37C; purified recombinant enzyme, pH 3.5, 37C
51.2
-
recombinant enzyme
56.4
-
native enzyme
2279
-
purified fraction from salivary gland carcinoma
84110
-
purified recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 4
recombinant enzyme, substrate 2-nitrophenyl 2-acetyl-2-deoxy-alpha-D-galactosamine
2.6 - 3.2
-
-
2.7 - 3.5
-
isoenzyme I, p-nitrophenyl-alpha-N-acetylgalactosaminide
3
-
isozyme alpha-GalNAcase II
3.5
-
isozyme alpha-GalNAcase I
3.9
-
forssman hapten
4 - 4.5
-
p-nitrophenyl-alpha-N-acetylgalactosaminide
4 - 6.8
-
-
4
-
p-nitrophenyl-alpha-N-acetylgalactosaminide
4.3 - 4.7
-
p-nitrophenyl-alpha-N-acetylgalactosaminide
4.3 - 4.7
-
p-nitrophenyl-alpha-N-acetylgalactosaminide
4.8
-
4-methylumbelliferyl-alpha-galactopyranoside
5.8 - 6.8
-
three isoforms
6.1
-
; enzyme from HIV patient serum
6.5 - 7
7 - 8
-
pH 7.2, 36C
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5 - 8
-
p-nitrophenyl-alpha-N-acetylgalactosaminide
3 - 8
-
below pH 3.6 the activity declines rapidly and at pH 3.0 retains about 1% of the orginal activity, above pH 7.5 the activity declines rapidly and at pH 8.0 retains about 4% of the orginal activity
4 - 7
-
not active above, p-nitrophenyl-alpha-N-acetylgalactosaminide
6 - 8
-
27% decline in activity from pH 6.0-8.0 with no apparent optimum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50 - 55
; recombinant enzyme, substrate 2-nitrophenyl 2-acetyl-2-deoxy-alpha-D-galactosamine
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.58
-
recombinant enzyme
7.1
-
purified isozyme alpha-GalNAcase II
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
inducible enzyme
Manually annotated by BRENDA team
-
high levels, salivary gland and gingival
Manually annotated by BRENDA team
-
299, low levels
Manually annotated by BRENDA team
-
low levels
Manually annotated by BRENDA team
-
cutaneous melanoma, specific enzymatic activity of serum alpha-N-acetylgalactosaminidase is significantly increased in stage III melanoma patients, but not in early stages
Manually annotated by BRENDA team
-
peripheral blood, inducible enzyme
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48000
-
gel filtration
70000
and 130000, gel filtration; recombinant enzyme, analytical ultracentrifugation and dynamic light scattering, monomeric form
80000
-
pH 4.2, gel filtration
84000
-
gel filtration
102000
-
gel filtration
115000
-
non-denaturating polyacrylamide gel electrophoresis
130000
and 70000, gel filtration; recombinant enzyme, analytical ultracentrifugation and dynamic light scattering, dimeric form
430000
-
about, isozyme alpha-GalNAcase I, gel filtration
440000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
homodimer
-
alpha-NAGAL is a homodimer with each monomer divided into two domains. Domain 1 forms a (beta/alpha)8 barrel, and domain 2 contains eight antiparallel beta strands in two beta sheets
monomer
multimer
polymer
-
x * 47000, isozyme alpha-GalNAcase I, SDS-PAGE
tetramer
-
4 * 50000, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme and in complex with alpha-N-acetylgalactosamine, to 2.3 and 2.4 A resolution, respectively. Each monomer of NagA consists of two closely associated domains, forming a narrow tunnel in which the NAD+ molecule is anchored. The NAD+ is embedded within a narrow tunnel, almost exclusively shielded from solvent. It is bound with its diphosphate group oriented towards the N-terminus of the first alpha-helix of the N-terminal dinucleotide-binding domain and has both the adenine ring and the nicotinamide ring in the anti conformation. No major structural changes are observed in NagA upon product binding
X-ray crystallography
-
purified recombinant wild-type and mutant enzymes in complexes with two catalytic products, the alpha-galactose and alpha-GalNAc monosaccharides, and a covalent intermediate bound in the enzyme active site, X-ray diffraction structure determination and analysis at 1.9 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.5 - 4
-
-
695797
2.3 - 6.5
-
purified isozyme alpha-GalNAcase II, stable at 4C
707791
2.5 - 8
-
-
208674
2.7 - 4.5
-
purified isozyme alpha-GalNAcase I, stable at 4C
707791
3 - 7.5
-
-
208650
3.5 - 8.5
-
purified enzyme, stable at 4C
707791
3.6 - 6.8
-
-
208661
4 - 6
-
-
208649
4.5 - 9.5
-
-
208673
4.5 - 5.5
-
isoenzyme I
208652
4.5 - 6.5
-
isoenzyme II
208652
5 - 8
-
-
657266
6 - 7.5
-
-
208648
additional information
-
comparison of activity at pH 5.0, 6.0 and 7.0 from Hep-G2 cells, HCT116 cells and Chang liver cells
655254
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 24
-
active and stable at
4 - 37
-
the enzyme hydrolyses A2 epitopes at 4C with 42% of the efficiency of that at 37C
35
-
stable for at least 3 days
42
-
comparison of stability at 42C of mutant and wild-type enzymes
45
-
55% loss of activity after 10 min
additional information
-
the enzyme does not withstand freezing
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 6 months
-20C, about 5% loss of activity
-
-20C, acidified plasma, at least one year
-
-20C, lyophilized, several years
0C, 1 month
-
0C, sodium citrate buffer, bovine serum albumin, 2 weeks, pH 4.2
-
20C, stable for 1 week
-
4C, 50 mM citrate-phosphate buffer within the pH range 2-4, stable for several weeks
4C, enzyme separation buffer containing 3 mM sodium azide, 25-31 months
-
4C, pH 1.5-4.0, stable for several months
-
4C, sterile condition, stable for any length of time
-
the purified wild-type and N201Q mutant proteins expressed in insect cells retain nearly full activity for months at 4C, but the CHO-expressed material loses most of its activity within 72 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme from salivary gland cells
-
enzyme mutant from cell culture by dialysis, ammonium sulfate fractionation, hydrophobic interaction, cation exchange, and anion exchange chromatography
-
native enzyme
-
native enzyme from cell culture supernatant by three chromatographic steps to homogeneity, recombinant enzyme from Saccharomyces cerevisiae to homogeneity 105.9fold by hydrophobic interaction chromatography, ultrafiltration, cation exchange chromatography, gel filtration, and anion exchange chromatography; recombinant protein
native enzyme from placenta by 4-aminophenyl thio-beta-D-galactose-ceramide hexoside-resin affinity
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native isozymes alpha-GalNAcase I and alpha-GalNAcase II from liver
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native isozymes alpha-GalNAcase I and II are isolated from liver homogenate by ammonium sulfate fractionation and separated from each other by gel filtration. Then they are further individually purified by cation exchange chromatography and gel filtration, and additionally by hydroxylapatite chromatography for isozyme alpha-GalNAcase II. Isozyme alpha-GalNAcase I is purified 440fold, isozyme alpha-GalNAcase II 544fold
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partially
recombinant enzyme
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recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) as inactive enzyme in inclusion bodies, expression in Pichia pastoris strain X33 is not successful, heterologous overexpression in Saccharomyces cerevisiae strain W303-1A without enzyme secretion; expression in Saccharomyces cerevisiae
expression in B-lymphoblast cell line
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expression in bacteria, rabbit reticulocyte lysate and yeast
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expression in Chines hamster ovary cells
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expression in CHO and COS-1 cells. Expression of wild-type and mutant enzymes in Escherichia coli as insoluble proteins, and in Kluyveromyces lactis as soluble hyperglycosylated proteins. Expression of wild-type and mutant enzymes in Trichoplusia ni Tn5 insect cells using the baculovirus transfection system leads to suitable proteins
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expression in Cos-1 cells
expression in Escherichia coli
expression in yeast Pichia pastoris
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expression of aagA gene in Escherichia coli, resulting in recombinants exhibiting high-level expression of the expected activity
expression of the engineered enzyme in CHO cells
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recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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the gene encoding the clostridial enzyme is cloned in an Escherichia coli T7 expression system
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D217N
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a naturally occuring Schindler disease and/or Kanzaki disease mutation
E193X
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a naturally occuring Schindler disease and/or Kanzaki disease mutation
E325K
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a naturally occuring Schindler disease and/or Kanzaki disease mutation
E367K
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a naturally occuring Schindler disease and/or Kanzaki disease mutation
N124Q
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35% activity in comparison to 100% activity of wild-type enzyme
N177Q
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48% activity in comparison to 100% activity of wild-type enzyme
N359Q
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57% activity in comparison to 100% activity of wild-type enzyme
N385Q
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4% activity in comparison to 100% activity of wild-type enzyme
S160C
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a naturally occuring Schindler disease and/or Kanzaki disease mutation
S188E/A191L
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Ser188 and Ala191 play important roles in the recognition of an N-acetylgalactosamine residue in NAGA, while lu203 and Leu206 play important roles in the recognition of a galactose residue in GLA. Construction of a modified alpha-N-acetylgalactosaminidase with alpha-galactosidase A-like substrate specificity. The enzyme acquires the ability to catalyze the degradation of 4-methylumbelliferyl-alpha-D-galactopyranoside, but retaines the wild-type NAGA's stability, overview
additional information
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construction of mutants of each of the five N-linked glycosylation sites
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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use of a modified alpha-N-acetylgalactosaminidase in the development of enzyme replacement therapy for Fabry disease
diagnostics
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the enzyme is used for blood group conversion together with alpha-galactosidase, best in glucose buffer
medicine
Show AA Sequence (106 entries)
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