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Literature summary extracted from

  • van Kasteren, P.B.; Bailey-Elkin, B.A.; James, T.W.; Ninaber, D.K.; Beugeling, C.; Khajehpour, M.; Snijder, E.J.; Mark, B.L.; Kikkert, M.
    Deubiquitinase function of arterivirus papain-like protease 2 suppresses the innate immune response in infected host cells (2013), Proc. Natl. Acad. Sci. USA, 110, E838-E847.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.B50 expressed in Escherichia coli BL21-Gold (DE3) cells equine arteritis virus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.22.B50 in complex with ubiquitin, hanging drop vapor diffusion method, using 100 mM MES, pH 6.2, 18% (w/v) polyethylene glycol 20,000 equine arteritis virus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.22.B50 C270A/H332A active site mutant equine arteritis virus
3.4.22.B50 I353R the mutant exhibits about 40fold reduction in specificity toward ubiquitin compared to the wild type while the activity toward RLRGG-7-amido-4-methylcoumarin is essentially unaltered equine arteritis virus
3.4.22.B50 I353W the mutant exhibits about 10fold reduction in specificity toward ubiquitin compared to the wild type while the activity toward RLRGG-7-amido-4-methylcoumarin is essentially unaltered equine arteritis virus
3.4.22.B50 T312A/I313V/I353R the mutant shows the greatest decrease in inhibitory activity in the interferon-beta promoter activity assay equine arteritis virus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.B50 Ub-3Br suicide inhibitor, i.e. ubiquitin(1-75)-3-bromopropylamide equine arteritis virus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.4.22.B50 ubiquitin + H2O equine arteritis virus
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.B50 equine arteritis virus P19811
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.B50 Ni-NTA column chromatography, Superdex 75 gel filtration, and Source 15Q column chromatography equine arteritis virus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.B50 RLRGG-7-amido-4-methylcoumarin + H2O
-
 equine arteritis virus RLRGG + 7-amino-4-methylcoumarin
-
 ?
3.4.22.B50 ubiquitin + H2O
-
 equine arteritis virus ?
-
 ?
3.4.22.B50 ubiquitin-aminomethylcoumarin + H2O
-
 equine arteritis virus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.B50 PLP2
-
 equine arteritis virus

General Information

EC Number General Information Comment Organism
3.4.22.B50 malfunction arteriviruses lacking PLP2 deubiquitinase activity elicit an enhanced host innate immune response equine arteritis virus
3.4.22.B50 physiological function the enzyme is essential for arterivirus replication by cleaving a site within the viral replicase polyproteins and also removes ubiquitin from cellular proteins. This deubiquitinase activity is a critical factor in arteriviral innate immune evasion equine arteritis virus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.22.B50 0.045
-
 RLRGG-7-amido-4-methylcoumarin wild type enzyme, at pH 8.0 and 37°C equine arteritis virus
3.4.22.B50 0.065
-
 RLRGG-7-amido-4-methylcoumarin mutant enzyme I353R, at pH 8.0 and 37°C equine arteritis virus
3.4.22.B50 0.155
-
 RLRGG-7-amido-4-methylcoumarin mutant enzyme I353W, at pH 8.0 and 37°C equine arteritis virus
3.4.22.B50 0.413
-
 ubiquitin-aminomethylcoumarin mutant enzyme I353R, at pH 8.0 and 37°C equine arteritis virus
3.4.22.B50 1.741
-
 ubiquitin-aminomethylcoumarin mutant enzyme I353W, at pH 8.0 and 37°C equine arteritis virus
3.4.22.B50 17
-
 ubiquitin-aminomethylcoumarin wild type enzyme, at pH 8.0 and 37°C equine arteritis virus