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Literature summary for 3.4.19.12 extracted from

  • Drag, M.; Mikolajczyk, J.; Bekes, M.; Reyes-Turcu, F.E.; Ellman, J.A.; Wilkinson, K.D.; Salvesen, G.S.
    Positional-scanning fluorigenic substrate libraries reveal unexpected specificity determinants of DUBs (deubiquitinating enzymes) (2008), Biochem. J., 415, 367-375.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information activation of the enzyme with increasing salt concentration, reaching a maximum above 0.8 M Severe acute respiratory syndrome-related coronavirus
Sodium citrate activation of the enzymes with increasing salt concentration, reaching a maximum above 0.8 M. Most effective activation in the case of UCH-L3, where 1.1 M sodium citrate enhances catalysis on the synthetic substrate Ac-LRGG-7-amido-4-trifluoromethylcoumarin about 24fold, compared with low-salt conditions. The tetrapeptide substrate Ac-LRGG-7-amido-4-trifluoromethylcoumarin and the pentapeptide substrate containing alanine at position P5, namely Ac-ALRGG-7-amido-4-trifluoromethylcoumarin, both demonstrate a salt effect, but the pentapeptide substrate containing arginine in position P5, Ac-RLRGG-7-amido-4-trifluoromethylcoumarin, does not demonstrate one Homo sapiens
Sodium citrate activation of the enzymes with increasing salt concentration, reaching a maximum above 0.8 M. The tetrapeptide substrate Ac-LRGG-7-amido-4-trifluoromethylcoumarin and the pentapeptide substrate containing alanine at position P5, namely Ac-ALRGG-7-amido-4-trifluoromethylcoumarin, both demonstrate a salt effect, but the pentapeptide substrate containing arginine in position P5, Ac-RLRGG-7-amido-4-trifluoromethylcoumarin, does not demonstrate one Homo sapiens
Sodium citrate activation of the enzymes with increasing salt concentration, reaching a maximum above 0.8 M. Weakest activation in the case of IsoT, where there is almost no increase in catalysis compared with low salt conditions. The tetrapeptide substrate Ac-LRGG-7-amido-4-trifluoromethylcoumarin and the pentapeptide substrate containing alanine at position P5, namely Ac-ALRGG-7-amido-4-trifluoromethylcoumarin, both demonstrate a salt effect, but the pentapeptide substrate containing arginine in position P5, Ac-RLRGG-7-amido-4-trifluoromethylcoumarin, does not demonstrate one Homo sapiens
ubiquitin 20fold enhancement of the cleavage of Ac-LRGG-7-amido-4-trifluoromethylcoumarin by IsoT in the presence of equimolar ubiquitin Homo sapiens
ubiquitin iIn the case of OTU-1 full-length ubiquitin does not result in any substantial decrease or increase of the processing of the fluorigenic substrate Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
UCH-L3 subcloned into a pET28 expression vector bearing an N-terminal histidine tag. Full-length UCH-L3 expressed in Escherichia coli BL21(DE3) cells Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
additional information very high sodium citrate concentrations reveal a decrease in activity; very high sodium citrate concentrations reveal a decrease in activity; very high sodium citrate concentrations reveal a decrease in activity, most dramatic for OTU-1, which is due to enzyme precipitation in the assay Homo sapiens
additional information very high sodium citrate concentrations reveal a decrease in activity. Full-length ubiquitin does not result in any substantial decrease or increase of the processing of the fluorigenic substrate Severe acute respiratory syndrome-related coronavirus
ubiquitin increasing ubiquitin concentrationin reveals a decrease in the cleavage efficiency, presumably as a result of competition between the second (substrate) ubiquitin molecule and the fluorigenic substrate binding to the enzyme active centre; increasing ubiquitin concentrationin reveals a decrease in the cleavage efficiency, presumably as a result of competition between the second (substrate) ubiquitin molecule and the fluorigenic substrate binding to the enzyme active centre. In the case of OTU-1 full-length ubiquitin does not result in any substantial decrease or increase of the processing of the fluorigenic substrate; increasing ubiquitin concentrationin reveals a decrease in the cleavage efficiency, presumably as a result of competition between the second (substrate) ubiquitin molecule and the fluorigenic substrate binding to the enzyme active centre. In the case of UCH-L3 inhibition of the fluorigenic substrate by elevated concentrations of full-length ubiquitin Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens P15374
-
-
Homo sapiens P45974
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-
Homo sapiens Q5VVQ6
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-
Severe acute respiratory syndrome-related coronavirus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
UCH-L3 purified on Ni-NTA column Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ac-ALRGG-7-amido-4-trifluoromethylcoumarin
-
Severe acute respiratory syndrome-related coronavirus ?
-
?
Ac-ALRGG-7-amido-4-trifluoromethylcoumarin
-
Homo sapiens ?
-
?
Ac-LRGG-7-amido-4-trifluoromethylcoumarin
-
Severe acute respiratory syndrome-related coronavirus ?
-
?
Ac-LRGG-7-amido-4-trifluoromethylcoumarin
-
Homo sapiens ?
-
?
Ac-RLRGG-7-amido-4-trifluoromethylcoumarin
-
Severe acute respiratory syndrome-related coronavirus ?
-
?
Ac-RLRGG-7-amido-4-trifluoromethylcoumarin
-
Homo sapiens ?
-
?
additional information Iso-T shows high preference for the natural Leu-Arg-Gly motif in the P4–P2 positions. Only reactivity with norleucine at P4 and lysine at P3 mar the exquisite specificity Homo sapiens ?
-
?
additional information OTU-1 shows similarly to UCH-L3 high specificity in the P2 and P3 positions and less specificity in the P4 position. The P4 position prefers norleucine over leucine, but other bulky amino acids are also accepted, such as tyrosine, tryptophan, phenylalanine or lysine Homo sapiens ?
-
?
additional information PLpro reveals very high specificity in the P4 position, where practically only leucine can be tolerated, and even higher specificity in the P2 position, with only glycine being accepted. However, the P3 position can accommodate a number of different amino acids, with some preference for the hydrophobes leucine and tyrosine, and also for the basic side chains of lysine and arginine, amino acids that possess a substantial hydrophobic character Severe acute respiratory syndrome-related coronavirus ?
-
?
additional information UCH-L3 shows a preference for leucine at P4, but also a broad tolerance of several other hydrophobic residues at this position. It also shows a surprising selectivity for alanine and valine at P2, in addition to the canonical glycine and high preference at the P3 position and relatively low specificity in the P4 position Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
Iso-T
-
Homo sapiens
isopeptidase T
-
Homo sapiens
OTU-1
-
Homo sapiens
ovarian tumour 1
-
Homo sapiens
papain-like protease
-
Severe acute respiratory syndrome-related coronavirus
PLpro
-
Severe acute respiratory syndrome-related coronavirus
ubiquitin C-terminal hydrolase L3
-
Homo sapiens
UCH-L3
-
Homo sapiens