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Literature summary for 3.2.1.31 extracted from

  • Islam, M.R.; Tomatsu, S.; Shah, G.N.; Grubb, J.H.; Jain, S.; Sly, W.S.
    Active site residue of human beta-glucuronidase. Evidence for Glu540 as the nucleophile and Glu451 as the acid-base residue (1999), J. Biol. Chem., 274, 23451-23455.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
azide 50 mM-0.5 M, stimulation of mutant enzyme E451A Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutant enzymes expressed COS cells and GUSB-deficient fibroblasts from the SV40 late promoter in vector pJC119 Homo sapiens

Protein Variants

Protein Variants Comment Organism
D207A 4.9% of the wild-type activity is expressed in COS cells Homo sapiens
E451A 0.6% of the wild-type activity is expressed in COS cells, tetrameric enzyme, ratio of turnover number to KM-value is decreased 9100fold as compared to wild-type enzyme. Mutant enzyme E541A is inactivated at a faster rate than the wild-type enzyme, and is completely inactivated within 30 min. 50 mM azide causes 70% loss of activity of wild-type enzyme and mutant enzyme E451A. 50 mM-0.5 M stimulates mutant enzyme E451A. 1 mM inhibits wild-type enzyme and mutant enzyme E451A Homo sapiens
E451Q 5.9% of the wild-type activity is expressed in COS cells Homo sapiens
E515A no activity is expressed in COS cells Homo sapiens
E540A no activity is expressed in COS cells, tetrameric enzyme, ratio of turnover number to KM-value is decreased 33000 fold as compared to wild-type enzyme, optimal pH is 5.0 instead of 4.5 for the wild-type enzyme. Mutant enzyme is inactivated at a faster rate than the wild-type enzyme at 68°C Homo sapiens
E540D no activity is expressed in COS cells Homo sapiens
E540Q 0.3% of the wild-type activity is expressed in COS cells Homo sapiens
Y504A 1.4% of the wild-type activity is expressed in COS cells, tetrameric enzyme, the ratio of turnover number to KM-value is decreased 830fold as compared to wild-type enzyme. Mutant enzyme is as stable as the wild-type enzyme at 68°C Homo sapiens
Y504F 2.3% of the wild-type activity is expressed in COS cells Homo sapiens
Y504H 0.6% of the wild-type activity is expressed in COS cells Homo sapiens
Y508A no activity is expressed in COS cells Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
azide 50 mM, 70% loss of activity of wild-type enzyme and mutant enzyme E451A. 50 mM-0.5 M, stimulation of mutant enzyme E451A. 1 mM, inhibition of wild-type enzyme and mutant enzyme E451A Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 5.0, mutant enzyme E540A Homo sapiens
1.33
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 4.0-5.0, mutant enzyme E451A Homo sapiens
1.67
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, mutant enzyme Y540A Homo sapiens
2.76
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 4.5, wild-type enzyme Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Homo sapiens
-
-

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
culture medium
-
Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl-beta-D-glucuronide + H2O
-
Homo sapiens 4-methylumbelliferone + D-glucuronic acid
-
?

Synonyms

Synonyms Comment Organism
hGUSB
-
Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
68
-
pH 7.5, 75 mM NaCL and 5 mg/ml bovine serum albumin, 2 h: less than 40% inactivation of wild-type enzyme, mutant enzyme Y504A is as stable as the wild-type enzyme, mutant enzyme E540A is inactivated at a faster rate than the wild-type enzyme, mutant enzyme E541A is completely inactivated within 30 min Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.004
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 5.0, mutant enzyme E540A Homo sapiens
0.015
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 4.0-5.0, mutant enzyme E451A Homo sapiens
0.2
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, mutant enzyme Y540A Homo sapiens
276
-
4-methylumbelliferyl-beta-D-glucuronide 37°C, pH 4.5, wild-type enzyme Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4 5 mutant enzyme E451A Homo sapiens
4.5
-
wild-type enzyme Homo sapiens
5
-
mutant enzyme E540A Homo sapiens