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
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
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
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.
ubiquitin + H2O
-
equine arteritis virus ?
-
?
RLRGG-7-amido-4-methylcoumarin + H2O
-
equine arteritis virus RLRGG + 7-amino-4-methylcoumarin
-
?
ubiquitin-aminomethylcoumarin + H2O
-
equine arteritis virus ?
-
?

Synonyms

Synonyms Comment Organism
PLP2
-
equine arteritis virus

General Information

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
17
-
ubiquitin-aminomethylcoumarin 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
0.045
-
RLRGG-7-amido-4-methylcoumarin wild type enzyme, 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