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Literature summary for 3.2.1.169 extracted from

  • Dong, D.L.; Hart, G.W.
    Purification and characterization of an O-GlcNAc selective N-acetyl-beta-D-glucosaminidase from rat spleen cytosol (1994), J. Biol. Chem., 269, 19321-19330.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information addition of Triton X-100 does not increase activity Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
2-(acetylamino)-2-deoxy-1-S-hexanoyl-1-thio-beta-D-glucopyranose inhibits activity at 10 mM, 47.5% relative activity compared with activity without any addition of effector Rattus norvegicus
2-acetamido-2-deoxy-D-gluconhydroxime-1,5-lactone inhibits activity at 0.1 mM, 6.7% relative activity compared with activity without any addition of effector Rattus norvegicus
2-acetamindo-1-amino-1,2-dideoxy-beta-D-glucopyranose inhibits activity at 10 mM, 3.2% relative activity compared with activity without any addition of effector Rattus norvegicus
2-acetamindo-1-azido-1,2-dideoxy-beta-D-glucopyranose inhibits activity at 10 mM, 19.3% relative activity compared with activity without any addition of effector Rattus norvegicus
2-acetamindo-1-bromo-1,2-dideoxy-beta-D-glucopyranose inhibits activity at 10 mM, 34.8% relative activity compared with activity without any addition of effector Rattus norvegicus
alpha-D-mannose inhibits activity at 10 mM, 62.6% relative activity compared with activity without any addition of effector Rattus norvegicus
castanospermine inhibits activity at 10 mM, 32.7% relative activity compared with activity without any addition of effector Rattus norvegicus
Cd2+ inhibits activity at 1 mM, 7.7% relative activity compared with activity without any addition of effector Rattus norvegicus
Cu2+ inhibits activity at 1 mM, 1% relative activity compared with activity without any addition of effector Rattus norvegicus
D-glucosamine inhibits activity at 10 mM, 60.7% relative activity compared with activity without any addition of effector Rattus norvegicus
deoxynojirimycin slight inhibition Rattus norvegicus
Hg2+ inhibits activity at 1 mM, 6% relative activity compared with activity without any addition of effector Rattus norvegicus
methyl alpha-D-mannose inhibits activity at 10 mM, 11.3% relative activity compared with activity without any addition of effector. Other sugars and sugar analogs do not inhibit with exception of methyl-alpha-D-mannose Rattus norvegicus
additional information addition of 50 mM GalNAc does not inhibit the neutral hexosaminidase activity; GlcNAc analogs with no or small effect on the enzyme activity: N-acetylgalactosamine, 4-aminophenyl 1-thio-beta-D-galactopyranoside, (2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl)-L-asparagine, 2-(acetylamino)-2-deoxy-N-hexanoyl-beta-D-glucopyranosylamine, 2-(acetylamino)-2-deoxy-1-O-phosphono-beta-D-glucopyranose, bromoconduritol, iodoacetic acid, N-acetylneuraminic acid Rattus norvegicus
N-acetyl-D-glucosamine inhibits activity at 10 mM, 40% relative activity compared with activity without any addition of effector Rattus norvegicus
N-ethylmaleimide inhibits activity at 10 mM, 15.3% relative activity compared with activity without any addition of effector Rattus norvegicus
N-methyl-deoxynojirimycin inhibits activity at 10 mM, 73.5% relative activity compared with activity without any addition of effector Rattus norvegicus
NaCl high salt, 1.0 M NaCl, inhibits enzyme activity by 50% compared with activity without any addition of effector Rattus norvegicus
O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate inhibits activity at 0.1 mM, 4.1% relative activity compared with activity without any addition of effector Rattus norvegicus
p-aminophenyl-S-GlcNAc inhibits activity at 10 mM, 76.3% relative activity compared with activity without any addition of effector Rattus norvegicus
Sn2+ inhibits activity at 1 mM, 57.6% relative activity compared with activity without any addition of effector Rattus norvegicus
Tris high salt, 75 mM, inhibits enzyme activity by 50% compared with activity without any addition of effector Rattus norvegicus
UDP-GlcNAc inhibits activity at 10 mM, 78.1% relative activity compared with activity without any addition of effector Rattus norvegicus
Zn2+ inhibits activity at 1 mM, 7.3% relative activity compared with activity without any addition of effector Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5 5 4-nitrophenyl-beta-D-GlcNAc
-
Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Rattus norvegicus 5829
-
nucleoplasm enriched in nuclei Rattus norvegicus 5654
-

Metals/Ions

Metals/Ions Comment Organism Structure
Cd2+ inhibits activity at 1 mM Rattus norvegicus
Cu2+ inhibits activity at 1 mM Rattus norvegicus
Hg2+ inhibits activity at 1 mM Rattus norvegicus
additional information EDTA, Mg2+, Fe3+, Ca2+, Mn2+, and Co2+ do not affect the activity Rattus norvegicus
Zn2+ inhibits activity at 1 mM Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
51000
-
1 * 54000 (alpha-subunit) and 1 * 51000 (beta-subunit) Rattus norvegicus
54000
-
1 * 54000 (alpha-subunit) and 1 * 51000 (beta-subunit) Rattus norvegicus
106000
-
SDS-PAGE and sucrose density gradient Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Rattus norvegicus strong selectivity for O-GlcNAc-synthetic glycopeptides. Enzyme does not cleave 4-nitrophenylGalNAc and is highly selective for O-GlcNAc bearing glycopeptides. O-GlcNAcase is involved in the regulated removal of O-GlcNAc from O-GlcNAc-bearing glycoproteins in the nucleoplasmic and cytoplasmic compartments of cells ?
-
?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
over 22000fold, gel filtration chromatography Rattus norvegicus

Source Tissue

Source Tissue Comment Organism Textmining
spleen enzyme is ubiquitously present in all rat tissues Rattus norvegicus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
last purification step: 13046.67 unit/mg. One unit is the amount of enzyme required to catalyze the release of 4-nitrophenyl from 1 mmol of 4-nitrophenylGlcNAc/min at 37°C Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-aminophenyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranoside + H2O 67% relative activity compared to 4-nitrophenyl-beta-D-GlcNac Rattus norvegicus ?
-
?
4-methylumbelliferyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranoside + H2O 33% relative activity compared to 4-nitrophenyl-beta-D-GlcNac Rattus norvegicus ?
-
?
4-nitrophenyl beta-D-GlcNAc + H2O
-
Rattus norvegicus 4-nitrophenol + N-acetyl-D-glucosamine
-
?
GlcNAc beta(1-4)GlcNAc + H2O 4% relative activity. Enzyme can cleave terminal Glc-NAc from disaccharides to a limited extent as long as GlcNAc is linked to the other saccharides beta-linkage at the C-1 position Rattus norvegicus N-acetyl-D-glucosamine + N-acetyl-D-glucosamine
-
?
GlcNAc beta(1-6)Gal + H2O 29% relative activity. Enzyme can cleave terminal Glc-NAc from disaccharides to a limited extent as long as GlcNAc is linked to the other saccharides beta-linkage at the C-1 position Rattus norvegicus N-acetyl-D-glucosamine + beta-D-galactose
-
?
GlcNAcbeta(1-6)Man-beta-1-OMe + H2O 33% relative activity. Enzyme can cleave terminal Glc-NAc from disaccharides to a limited extent as long as GlcNAc is linked to the other saccharides beta-linkage at the C-1 position Rattus norvegicus N-acetyl-D-glucosamine + methyl beta-D-mannoside
-
?
additional information strong selectivity for O-GlcNAc-synthetic glycopeptides. Enzyme does not cleave 4-nitrophenylGalNAc and is highly selective for O-GlcNAc bearing glycopeptides. O-GlcNAcase is involved in the regulated removal of O-GlcNAc from O-GlcNAc-bearing glycoproteins in the nucleoplasmic and cytoplasmic compartments of cells Rattus norvegicus ?
-
?
additional information shows no activity against other monosaccharides either in alpha or beta linkage to 4-nitrophenyl, such as, 4-nitrophenyl-alpha-D-GlcNAc, 4-nitrophenyl-alpha-D-glucose, 4-nitrophenyl-beta-D-glucose, 4-nitrophenyl-alpha-D-GalNAc, 4-nitrophenyl-beta-D-GalNAc, 4-nitrophenyl-alpha-D-galactose, 4-nitrophenyl-beta-D-galactose, 4-nitrophenyl-alpha-D-mannose, 4-nitrophenyl-alpha-L-fructose Rattus norvegicus ?
-
?
additional information substitution of GlcNAc at other positions prevents cleavage, for example, Gal(1-3)GlcNAc-p-nitrophenol, Gal(1-4)GlcNAc, Gal(1-3)GlcNAc, and UDP-GlcNAc, the enzyme specifically cleave GlcNAc at C-1 position in beta-linkage to other groups Rattus norvegicus ?
-
?
P-(GlcNAc)STST + H2O
-
Rattus norvegicus PSTST + N-acetyl-D-glucosamine
-
?
YSD-(GlcNAc)SPSTST + H2O
-
Rattus norvegicus YSDSPSTST + N-acetyl-D-glucosamine
-
?

Subunits

Subunits Comment Organism
heterodimer 1 * 54000 (alpha-subunit) and 1 * 51000 (beta-subunit) Rattus norvegicus

Synonyms

Synonyms Comment Organism
cytoplasmic O-GlcNAcase
-
Rattus norvegicus
neutral O-GlcNAcase
-
Rattus norvegicus
O-GlcNAc selective N-acetyl-beta-D-glucosaminidase
-
Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at and most active at Rattus norvegicus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4 37 most active at 37°C, and less active at room temperature, and very little activity at 4°C Rattus norvegicus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
20 37 enzyme loses activity very fast (within 5 min) at 57°C, stable at 37°C and room temperature Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.4
-
at pH below 5.0, it begins to lose activity dramatically, at pH 6 or higher, it maintains high activity Rattus norvegicus

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5
-
enzyme is unstable below pH 4.5 Rattus norvegicus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000052
-
O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
-
Rattus norvegicus
0.00168
-
2-acetamido-2-deoxy-D-gluconhydroxime-1,5-lactone
-
Rattus norvegicus
0.151
-
2-acetamindo-1-amino-1,2-dideoxy-beta-D-glucopyranose
-
Rattus norvegicus
1.14
-
2-acetamindo-1-azido-1,2-dideoxy-beta-D-glucopyranose
-
Rattus norvegicus
3.9
-
N-acetyl-D-glucosamine
-
Rattus norvegicus

pI Value

Organism Comment pI Value Maximum pI Value
Rattus norvegicus
-
-
5.7

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00015
-
-
Rattus norvegicus O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
0.007
-
-
Rattus norvegicus 2-acetamido-2-deoxy-D-gluconhydroxime-1,5-lactone
0.16
-
-
Rattus norvegicus 2-acetamindo-1-amino-1,2-dideoxy-beta-D-glucopyranose
1.5
-
-
Rattus norvegicus 2-acetamindo-1-azido-1,2-dideoxy-beta-D-glucopyranose
7
-
-
Rattus norvegicus N-acetyl-D-glucosamine
30
-
-
Rattus norvegicus D-glucosamine