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Literature summary extracted from

  • Larsen, C.N.; Price, J.S.; Wilkinson, K.D.
    Substrate binding and catalysis by ubiquitin C-terminal hydrolases: identification of two active site residues (1996), Biochemistry, 35, 6735-6744.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.19.12 overexpression of wild-type and mutant isozymes L1 and L3 in Escherichia coli BL21(DE3) Homo sapiens
3.4.19.12 UCH-L1, UCH-L3 Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.4.19.12 C90S site-directed mutagenesis, isozyme L1, inactive Homo sapiens
3.4.19.12 D176N site-directed mutagenesis, isozyme L1, 97.5% reduced activity compared to wild-type Homo sapiens
3.4.19.12 H161D site-directed mutagenesis, isozyme L1, very low activity Homo sapiens
3.4.19.12 H161K catalytically inactive Homo sapiens
3.4.19.12 H161K site-directed mutagenesis, isozyme L1, nearly inactive Homo sapiens
3.4.19.12 H161N catalytically inactive Homo sapiens
3.4.19.12 H161N site-directed mutagenesis, isozyme L1, nearly inactive Homo sapiens
3.4.19.12 H161Q catalytically inactive Homo sapiens
3.4.19.12 H161Q site-directed mutagenesis, isozyme L1, nearly inactive Homo sapiens
3.4.19.12 H161Y catalytically inactive Homo sapiens
3.4.19.12 H161Y site-directed mutagenesis, isozyme L1, nearly inactive Homo sapiens
3.4.19.12 H97N 85% activity compared to wild-type Homo sapiens
3.4.19.12 H97N site-directed mutagenesis, isozyme L1, slightly reduced activity Homo sapiens
3.4.19.12 H97Q 85% activity compared to wild-type Homo sapiens
3.4.19.12 H97Q site-directed mutagenesis, isozyme L1, slightly reduced activity Homo sapiens
3.4.19.12 Q37R site-directed mutagenesis, isozyme L1, unaltered activity Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.19.12 additional information
-
additional information
-
Bos taurus
3.4.19.12 additional information
-
additional information substrate-binding affinity of isozyme L3 Homo sapiens
3.4.19.12 0.0006
-
ubiquitin ethyl ester
-
Bos taurus
3.4.19.12 0.0006
-
ubiquitin ethyl ester isozyme L1 Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.4.19.12 Bos taurus
-
calf thymus
-
3.4.19.12 Bos taurus
-
isozyme L3, adult brain
-
3.4.19.12 Homo sapiens
-
-
-
3.4.19.12 Homo sapiens
-
isozymes L1, L2, and L3
-
3.4.19.12 Mammalia
-
-
-
3.4.19.12 Rattus norvegicus
-
isozyme L1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.19.12 recombinant wild-type and mutants of isozymes L1 and L3 from Escherichia coli BL21(DE3), to near homogeneity Homo sapiens
3.4.19.12 UCH-L1, UCH-L3 Homo sapiens

Reaction

EC Number Reaction Comment Organism Reaction ID
3.4.19.12 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

EC Number Source Tissue Comment Organism Textmining
3.4.19.12 brain
-
Homo sapiens
-
3.4.19.12 brain
-
Rattus norvegicus
-
3.4.19.12 brain isozyme L1, adult Bos taurus
-
3.4.19.12 neuron isozyme L1 Homo sapiens
-
3.4.19.12 thymus calf Bos taurus
-
3.4.19.12 thymus isozyme L3 Homo sapiens
-
3.4.19.12 thymus isozyme L3 Bos taurus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.19.12 25
-
-
Homo sapiens
3.4.19.12 25
-
purified isozyme L3, adult brain, substrate ubiquitin ethyl ester Bos taurus
3.4.19.12 25
-
purified wild-type isozyme L1, substrate ubiquitin ethyl ester Bos taurus
3.4.19.12 30
-
UCH-L1, ubiquitin ethyl ester as substrate Homo sapiens
3.4.19.12 30
-
purified recombinant isozyme L1, substrate ubiquitin ethyl ester Homo sapiens
3.4.19.12 110
-
UCH-L3, ubiquitin ethyl ester as substrate Homo sapiens
3.4.19.12 110
-
purified recombinant isozyme L3, substrate ubiquitin ethyl ester Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.19.12 ubiquitin ethyl ester + H2O
-
Homo sapiens ubiquitin + ethanol
-
?
3.4.19.12 ubiquitin ethyl ester + H2O
-
Mammalia ubiquitin + ethanol
-
?
3.4.19.12 ubiquitin ethyl ester + H2O
-
Bos taurus ubiquitin + ethanol
-
?

Synonyms

EC Number Synonyms Comment Organism
3.4.19.12 UCH
-
Homo sapiens
3.4.19.12 UCH
-
Bos taurus
3.4.19.12 UCH
-
Rattus norvegicus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.19.12 52
-
thermodynamic of denaturation, enthalpy, entropy, Gibbs energy Homo sapiens
3.4.19.12 52
-
reversible denaturation of wild-type isozymes L1 and L3, and of L1 mutants C90S, H161D, H161K, H161Y, thermodynamics Homo sapiens

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.4.19.12 Mammalia about
-
5