Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21-Gold (DE3) cells | equine arteritis virus |
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 | 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 | Comment | Organism | Structure |
---|---|---|---|
Ub-3Br | suicide inhibitor, i.e. ubiquitin(1-75)-3-bromopropylamide | equine arteritis virus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ubiquitin + H2O | equine arteritis virus | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
equine arteritis virus | P19811 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography, Superdex 75 gel filtration, and Source 15Q column chromatography | equine arteritis virus |
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 | Comment | Organism |
---|---|---|
PLP2 | - |
equine arteritis virus |
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 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 |