OTUB2 [GST-tagged]


Catalogue Number
64-0012-050
Product Size
50 µg
Price £
£250
Accession Number
NP_075601.1
Residues Expressed
1-234
Certificate of Analysis Size
50 µg
Species
Human
Source
E. coli expression
Quantity
50 µg
Storage
-70°C
Concentration
0.5 mg/ml
Formulation
50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol
Molecular Weight
~54 kDa
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: NP_075601.1. For full protein sequence information download the Certificate of Analysis pdf.
QA; Protein Identification
Confirmed by mass spectrometry.
QA Activity

Deubiquitylating Enzyme Assay: The activity of GST-OTUB2 was validated by determining the increase in fluorescence measured as a result of the enzyme catalysed cleavage of the fluorogenic substrate Ubiquitin-Rhodamine110-Glycine generating Ubiquitin and Rhodamine110-Glycine. Incubation of the substrate in the presence or absence of GST-OTUB2 was compared confirming the deubiquitylating activity of GST-OTUB2.


Background

The deubiquitylating enzymes (DUBs) regulate ubiquitin dependent signaling pathways. The activities of the DUBs include the generation of free ubiquitin from precursor molecules, the recycling of ubiquitin following substrate degradation to maintain cellular ubiquitin homeostasis and the removal of ubiquitin or ubiquitin-like protein (UBL) modifications through chain editing to rescue proteins from proteasomal degradation or to influence cell signalling events (Komander et al., 2009). There are two main classes of DUB enzyme the cysteine proteases and metalloproteases. OTUB2 is a cysteine protease and member of the Ovarian Tumor (OTU) protein super family. Cloning of the human OTUB2 gene was first described by Balakirev et al., (2003). OTUB2 contains Ubiquitin Interaction Motifs (UIMs) and Ubiquitin Associated (UBA) domains, as well as putative nuclear localization signals and a consensus LxxLL motif (Balakirev et al., 2003). De-ubiquitylation has emerged as a post translational mechanism for the activation of virus triggered type I interferon (IFN) (2) induction pathways. OTUB2 has been found to negatively regulate virus triggered type I IFN activation of IRF3 and NF-kappaB by mediating the de-ubiquitylation of TRAF3 and TRAF6, two ubiquitin E3 ligases involved in virus-triggered IRF3 and NF-kappaB activation pathways (Li et al., 2010). Integrative gene tissue array analysis has been used to locate disease relevant proteins to a linkage region associated with familial Amyotrophic Lateral Sclerosis (ALS)/frontotemporal dementia. Using this approach OTUB2 was identified as a disease relevant protein involved in TAU-1 and SOD1 induced motor neuron degeneration found in human sporadic ALS (SALS) (Kudo et al., 2010).


References

Balakirev MY, Tcherniuk SO, Jaquinod M, Chroboczek J (2003) Otubains: a new

family of cysteine proteases in the ubiquitin pathway. EMBO Rep 4, 517-22.

Komander D, Clague MJ, Urbe S (2009) Breaking the chains: structure and function

of the deubiquitinases. Nat Rev Mol Cell Biol 10, 550-63.

Kudo LC, Parfenova L, Vi N, Lau K, Pomakian J, Valdmanis P, Rouleau GA, Vinters HV, Wiedau-Pazos M, Karsten S (2010) Integrative gene-tissue microarray-based approach for identification of human disease biomarkers: application to amyotrophic lateral sclerosis. Hum Mol Genet 19, 3233 - 3253.

Li S, Zheng H, Mao AP, Zhong B, Li Y, Liu Y, Gao Y, Ran Y, Tien P, Shu HB (2010) Regulation of virus-triggered signaling by OTUB1- and OTUB2-mediated deubiquitination of TRAF3 and TRAF6. J Biol Chem 285, 4291-7.