Cloned (Comment) | Organism |
---|---|
expression of GST-tagged wild-type UCHL1 and mutant S18Y in Escherichia coli strain BL21(Rosetta) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type UCH-L1 and the Parkinson disease-associated variant of the enzyme, mutant S18Y, bound to a ubiquitin-based suicide substrate, ubiquitin vinyl methyl ester, hanging drop vapour diffusion method, 25 mg/ml enzyme-UbVMe complex in 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, and 10 mM DTT, are mixed with 2.4 M ammonium sulfate and 0.1 M bicine, pH 9.0, 2 months, X-ray diffraction structure determination and analysis at 2.4-2.85 A resolution, molecular replacement | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C90S | a catalytically inactive mutant of UCHL1 | Homo sapiens |
F214A | the F214A mutant binds with approximately 60fold less affinity to ubiquitin compared to the wild-type UCH-L1, it shows highly reduced activity compared tot he wild-type enzyme | Homo sapiens |
I93M | the mutant is a Parkinson disease-associated variant of UCH-L1, structure determination and comparison | Homo sapiens |
S18Y | the mutant is a Parkinson disease-associated variant of UCH-L1, structure determination and comparison | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ubiquitin vinyl methyl ester | a ubiquitin-based suicide substrate, binding structure analysis with wild-type and mutant S18Y enzymes, overview | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P09936 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged wild-type UCHL1 and mutant S18Y from Escherichia coli strain BL21(Rosetta) by glutathione affinity chromatography, cleavage of the tag, and gel filtration | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | - |
Homo sapiens | - |
neuron | UCHL1 is abundantly and selectively expressed in neurons | Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Phe214 is essential for distal-site substrate binding, and the loss of catalytic activity of the mutant may reflect its reduced affinity toward the substrate. The crystal structure of apo UCHL1 shows that the active-site residues are not aligned in a canonical form, with the nucleophilic cysteine being 7.7 A from the general base histidine, an arrangement consistent with an inactive form of the enzyme | Homo sapiens | ? | - |
? | |
ubiquitin-4-methylcoumarin 7-amide + H2O | - |
Homo sapiens | ubiquitin + 7-amino-4-methylcoumarin | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the crystal structure of apo UCHL1 shows that the active-site residues are not aligned in a canonical form, with the nucleophilic cysteine being 7.7 A from the general base histidine, an arrangement consistent with an inactive form of the enzyme | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
ubiquitin carboxy-terminal hydrolase L1 | - |
Homo sapiens |
ubiquitin hydrolase | - |
Homo sapiens |
UCH-L1 | - |
Homo sapiens |
UCHL1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
additional information | UCHL1 is a Parkinson disease-associated, putative cysteine protease | Homo sapiens |