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Literature summary for 3.4.19.12 extracted from

  • Gabriel, J.M.; Lacombe, T.; Carobbio, S.; Paquet, N.; Bisig, R.; Cox, J.A.; Jaton, J.C.
    Zinc is required for the catalytic activity of the human deubiquitinating isopeptidase T (2002), Biochemistry, 41, 13755-13766.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
isozymes ISOT-S and ISOT-L, cloning from brain mRNA, expression of both isozyme wild-types and mutant isozyme ISOT-S C335S in Escherichia coli strain MC1061 Homo sapiens

Protein Variants

Protein Variants Comment Organism
C335S site-directed mutagenesis of active site Cys335, unaltered substrate binding Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
dipicolinic acid
-
Homo sapiens
DTT erythrocyte isozyme ISOT-S and ISOT-L, inhibition by chelating of Zn2+ Homo sapiens
EDTA
-
Homo sapiens
EGTA
-
Homo sapiens
nitrilo-triacetate
-
Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ 0.2 mol per mol of protein, native isozyme ISOT-S Homo sapiens
Ni2+ 0.5 mol per mol of protein, native isozyme ISOT-S Homo sapiens
Zn2+ the Zn2+ bound to binding site 1 can be removed slowly by dialysis against 1,10-phenanthroline at pH 5.5, or rapidly by treatment at pH 3.0 in presence of 6 M urea followed by gel filtration at neutral pH Homo sapiens
Zn2+ removal and reconstitution methods, overview Homo sapiens
Zn2+ 0.9 mol per mol of protein, native isozyme ISOT-S Homo sapiens
Zn2+ both isozymes contain each two Zn2+-binding sites: one essential for proteolytic activity with high affinity as part of a cryptic nitrilo-triacetate-resistant pocket, a second which is enclosed with an excess of Zn2+ Homo sapiens
Zn2+ isozyme ISOT-S: Ni2+ can substitute by 90% for Zn2+, Cu2+ by 10% Homo sapiens
Zn2+ absolutely required, apoenzyme is inactive Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
isozymes ISOT-S and ISOT-L
-

Purification (Commentary)

Purification (Comment) Organism
isozyme ISOT-S from erythrocytes Homo sapiens
partial, recombinant wild-type and mutant isozymes ISOT-S and ISOT-L from Escherichia coli Homo sapiens

Renatured (Commentary)

Renatured (Comment) Organism
reconstitution of the apoenzyme with Zn2+ at pH 5.5 under nondenaturing conditions completely restores activity Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
brain isozymes ISOT-S and ISOT-L Homo sapiens
-
erythrocyte native isozyme ISOT-S Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
branched di-ubiquitin + H2O recombinant substrate expressed in Escherichia coli Homo sapiens ubiquitin
-
?
di-ubiquitin + H2O recombinant linear substrate, expressed in Escherichia coli Homo sapiens ubiquitin
-
?

Synonyms

Synonyms Comment Organism
deubiquinating isopeptidase T
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Homo sapiens