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 | Comment | Organism |
---|---|---|
C335S | site-directed mutagenesis of active site Cys335, unaltered substrate binding | Homo sapiens |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
isozymes ISOT-S and ISOT-L | - |
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 (Comment) | Organism |
---|---|
reconstitution of the apoenzyme with Zn2+ at pH 5.5 under nondenaturing conditions completely restores activity | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | isozymes ISOT-S and ISOT-L | Homo sapiens | - |
erythrocyte | native isozyme ISOT-S | Homo sapiens | - |
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 | Comment | Organism |
---|---|---|
deubiquinating isopeptidase T | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Homo sapiens |