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Literature summary for 3.4.22.B50 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)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-Gold (DE3) cells equine arteritis virus

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
C270A/H332A active site mutant equine arteritis virus
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
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
T312A/I313V/I353R the mutant shows the greatest decrease in inhibitory activity in the interferon-beta promoter activity assay equine arteritis virus

Inhibitors

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
equine arteritis virus P19811
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
PLP2
-
equine arteritis virus

General Information

General Information Comment Organism
malfunction arteriviruses lacking PLP2 deubiquitinase activity elicit an enhanced host innate immune response equine arteritis virus
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]

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