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Literature summary for 3.4.19.12 extracted from
- Substrate binding and catalysis by ubiquitin C-terminal hydrolases: identification of two active site residues (1996), Biochemistry, 35, 6735-6744.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpression of wild-type and mutant isozymes L1 and L3 in Escherichia coli BL21(DE3) | Homo sapiens |
| UCH-L1, UCH-L3 | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C90S | site-directed mutagenesis, isozyme L1, inactive | Homo sapiens |
| D176N | site-directed mutagenesis, isozyme L1, 97.5% reduced activity compared to wild-type | Homo sapiens |
| H161D | site-directed mutagenesis, isozyme L1, very low activity | Homo sapiens |
| H161K | catalytically inactive | Homo sapiens |
| H161K | site-directed mutagenesis, isozyme L1, nearly inactive | Homo sapiens |
| H161N | catalytically inactive | Homo sapiens |
| H161N | site-directed mutagenesis, isozyme L1, nearly inactive | Homo sapiens |
| H161Q | catalytically inactive | Homo sapiens |
| H161Q | site-directed mutagenesis, isozyme L1, nearly inactive | Homo sapiens |
| H161Y | catalytically inactive | Homo sapiens |
| H161Y | site-directed mutagenesis, isozyme L1, nearly inactive | Homo sapiens |
| H97N | 85% activity compared to wild-type | Homo sapiens |
| H97N | site-directed mutagenesis, isozyme L1, slightly reduced activity | Homo sapiens |
| H97Q | 85% activity compared to wild-type | Homo sapiens |
| H97Q | site-directed mutagenesis, isozyme L1, slightly reduced activity | Homo sapiens |
| Q37R | site-directed mutagenesis, isozyme L1, unaltered activity | Homo sapiens |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | - |
Bos taurus | |
| additional information | - |
additional information | substrate-binding affinity of isozyme L3 | Homo sapiens | |
| 0.0006 | - |
ubiquitin ethyl ester | - |
Bos taurus | |
| 0.0006 | - |
ubiquitin ethyl ester | isozyme L1 | Homo sapiens |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bos taurus | - |
calf thymus | - |
| Bos taurus | - |
isozyme L3, adult brain | - |
| Homo sapiens | - |
- |
- |
| Homo sapiens | - |
isozymes L1, L2, and L3 | - |
| Mammalia | - |
- |
- |
| Rattus norvegicus | - |
isozyme L1 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutants of isozymes L1 and L3 from Escherichia coli BL21(DE3), to near homogeneity | Homo sapiens |
| UCH-L1, UCH-L3 | Homo sapiens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| Thiol-dependent hydrolysis of ester, thioester, amide, peptide and isopeptide bonds formed by the C-terminal Gly of ubiquitin (a 76-residue protein attached to proteins as an intracellular targeting signal) | Cys90 and His161 are involved in catalytic rate enhancement | Homo sapiens |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| brain | - |
Homo sapiens | - |
| brain | - |
Rattus norvegicus | - |
| brain | isozyme L1, adult | Bos taurus | - |
| neuron | isozyme L1 | Homo sapiens | - |
| thymus | calf | Bos taurus | - |
| thymus | isozyme L3 | Homo sapiens | - |
| thymus | isozyme L3 | Bos taurus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 25 | - |
- |
Homo sapiens |
| 25 | - |
purified isozyme L3, adult brain, substrate ubiquitin ethyl ester | Bos taurus |
| 25 | - |
purified wild-type isozyme L1, substrate ubiquitin ethyl ester | Bos taurus |
| 30 | - |
UCH-L1, ubiquitin ethyl ester as substrate | Homo sapiens |
| 30 | - |
purified recombinant isozyme L1, substrate ubiquitin ethyl ester | Homo sapiens |
| 110 | - |
UCH-L3, ubiquitin ethyl ester as substrate | Homo sapiens |
| 110 | - |
purified recombinant isozyme L3, substrate ubiquitin ethyl ester | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ubiquitin ethyl ester + H2O | - |
Mammalia | ubiquitin + ethanol | - |
? |  |
| ubiquitin ethyl ester + H2O | - |
Homo sapiens | ubiquitin + ethanol | - |
? | |
| ubiquitin ethyl ester + H2O | - |
Bos taurus | ubiquitin + ethanol | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| UCH | - |
Homo sapiens |
| UCH | - |
Rattus norvegicus |
| UCH | - |
Bos taurus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 52 | - |
thermodynamic of denaturation, enthalpy, entropy, Gibbs energy | Homo sapiens |
| 52 | - |
reversible denaturation of wild-type isozymes L1 and L3, and of L1 mutants C90S, H161D, H161K, H161Y, thermodynamics | Homo sapiens |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Mammalia | about | - |
5 |
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