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Literature summary for 3.4.19.12 extracted from

  • Boudreaux, D.A.; Maiti, T.K.; Davies, C.W.; Das, C.
    Ubiquitin vinyl methyl ester binding orients the misaligned active site of the ubiquitin hydrolase UCHL1 into productive conformation (2010), Proc. Natl. Acad. Sci. USA, 107, 9117-9122.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of GST-tagged wild-type UCHL1 and mutant S18Y in Escherichia coli strain BL21(Rosetta) Homo sapiens

Crystallization (Commentary)

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

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

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

Organism UniProt Comment Textmining
Homo sapiens P09936
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
brain
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Homo sapiens
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neuron UCHL1 is abundantly and selectively expressed in neurons Homo sapiens
-

Substrates and Products (Substrate)

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
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Homo sapiens ubiquitin + 7-amino-4-methylcoumarin
-
?

Subunits

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

Synonyms Comment Organism
ubiquitin carboxy-terminal hydrolase L1
-
Homo sapiens
ubiquitin hydrolase
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Homo sapiens
UCH-L1
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Homo sapiens
UCHL1
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Homo sapiens

General Information

General Information Comment Organism
additional information UCHL1 is a Parkinson disease-associated, putative cysteine protease Homo sapiens