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Literature summary 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

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.169 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
3.2.1.169 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
3.2.1.169 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
3.2.1.169 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
3.2.1.169 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
3.2.1.169 alpha-D-mannose inhibits activity at 10 mM, 62.6% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 castanospermine inhibits activity at 10 mM, 32.7% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 Cd2+ inhibits activity at 1 mM, 7.7% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 Cu2+ inhibits activity at 1 mM, 1% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 D-glucosamine inhibits activity at 10 mM, 60.7% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 deoxynojirimycin slight inhibition Rattus norvegicus
3.2.1.169 Hg2+ inhibits activity at 1 mM, 6% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 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
3.2.1.169 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
3.2.1.169 N-acetyl-D-glucosamine inhibits activity at 10 mM, 40% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 N-ethylmaleimide inhibits activity at 10 mM, 15.3% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 N-methyl-deoxynojirimycin inhibits activity at 10 mM, 73.5% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 NaCl high salt, 1.0 M NaCl, inhibits enzyme activity by 50% compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 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
3.2.1.169 p-aminophenyl-S-GlcNAc inhibits activity at 10 mM, 76.3% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 Sn2+ inhibits activity at 1 mM, 57.6% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 Tris high salt, 75 mM, inhibits enzyme activity by 50% compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 UDP-GlcNAc inhibits activity at 10 mM, 78.1% relative activity compared with activity without any addition of effector Rattus norvegicus
3.2.1.169 Zn2+ inhibits activity at 1 mM, 7.3% relative activity compared with activity without any addition of effector Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.169 2.5 5 4-nitrophenyl-beta-D-GlcNAc
-
 Rattus norvegicus

Localization

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

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.169 51000
-
 1 * 54000 (alpha-subunit) and 1 * 51000 (beta-subunit) Rattus norvegicus
3.2.1.169 54000
-
 1 * 54000 (alpha-subunit) and 1 * 51000 (beta-subunit) Rattus norvegicus
3.2.1.169 106000
-
 SDS-PAGE and sucrose density gradient Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.169 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

EC Number Organism UniProt Comment Textmining
3.2.1.169 Rattus norvegicus
-
-
-

Purification (Commentary)

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

Source Tissue

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.169 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.169 4-aminophenyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranoside + H2O 67% relative activity compared to 4-nitrophenyl-beta-D-GlcNac Rattus norvegicus ?
-
 ?
3.2.1.169 4-methylumbelliferyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranoside + H2O 33% relative activity compared to 4-nitrophenyl-beta-D-GlcNac Rattus norvegicus ?
-
 ?
3.2.1.169 4-nitrophenyl beta-D-GlcNAc + H2O
-
 Rattus norvegicus 4-nitrophenol + N-acetyl-D-glucosamine
-
 ?
3.2.1.169 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
-
 ?
3.2.1.169 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
-
 ?
3.2.1.169 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
-
 ?
3.2.1.169 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 ?
-
 ?
3.2.1.169 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 ?
-
 ?
3.2.1.169 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 ?
-
 ?
3.2.1.169 P-(GlcNAc)STST + H2O
-
 Rattus norvegicus PSTST + N-acetyl-D-glucosamine
-
 ?
3.2.1.169 YSD-(GlcNAc)SPSTST + H2O
-
 Rattus norvegicus YSDSPSTST + N-acetyl-D-glucosamine
-
 ?

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.169 37
-
 assay at and most active at Rattus norvegicus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.169 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.169 20 37 enzyme loses activity very fast (within 5 min) at 57°C, stable at 37°C and room temperature Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.169 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

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.169 4.5
-
 enzyme is unstable below pH 4.5 Rattus norvegicus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.169 0.000052
-
 O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
-
 Rattus norvegicus
3.2.1.169 0.00168
-
 2-acetamido-2-deoxy-D-gluconhydroxime-1,5-lactone
-
 Rattus norvegicus
3.2.1.169 0.151
-
 2-acetamindo-1-amino-1,2-dideoxy-beta-D-glucopyranose
-
 Rattus norvegicus
3.2.1.169 1.14
-
 2-acetamindo-1-azido-1,2-dideoxy-beta-D-glucopyranose
-
 Rattus norvegicus
3.2.1.169 3.9
-
 N-acetyl-D-glucosamine
-
 Rattus norvegicus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.169 Rattus norvegicus
-
-
 5.7

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.2.1.169 0.00015
-
-
 Rattus norvegicus O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
3.2.1.169 0.007
-
-
 Rattus norvegicus 2-acetamido-2-deoxy-D-gluconhydroxime-1,5-lactone
3.2.1.169 0.16
-
-
 Rattus norvegicus 2-acetamindo-1-amino-1,2-dideoxy-beta-D-glucopyranose
3.2.1.169 1.5
-
-
 Rattus norvegicus 2-acetamindo-1-azido-1,2-dideoxy-beta-D-glucopyranose
3.2.1.169 7
-
-
 Rattus norvegicus N-acetyl-D-glucosamine
3.2.1.169 30
-
-
 Rattus norvegicus D-glucosamine