USP20 [GST-tagged]


Catalogue Number
64-0039-050
Product Size
50 µg
Price £
£430
Accession Number
AAH39593
Residues Expressed
1-913
Certificate of Analysis Size
50 µg
Species
human
Source
Insect sf21
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
128.7
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: AAH39593. For full protein sequence information download the Certificate of Analysis pdf.
QA; Protein Identification
Confirmed by mass spectrometry.
QA Activity

Deubiquitylase Enzyme Assay: The activity of GST-USP20 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-USP20 was compared confirming the deubiquitylating activity of GST-USP20.


Background

Deconjugating enzymes (DCEs) are proteases that process ubiquitin or ubiquitin-like gene products, reverse the modification of proteins by a single ubiquitin or ubiquitin-like protein (UBL) and remodel polyubiquitin (or poly-UBL) chains on target proteins (Reyes-Turcu et al., 2009). The deubiquitylating - or deubiquitinating - enzymes (DUBs) represent the largest family of DCEs and regulate ubiquitin dependent signalling 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 proteins (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, cysteine proteases and metalloproteases. Ubiquitin specific protease 20 (USP20) is a member of the cysteine protease enzyme family and cloning of the gene was first described by Nagase et al. (1999). USP20 has been shown to specifically deubiquitylate and stabilise Hypoxia-inducible factor (HIF)-1α, increasing expression of HIF-1α targeted genes, such as vascular endothelial growth factor (VEGF) (Li et al., 2005). USP20 also deubiquitylates tumour necrosis receptor-associated factor 6 (TRAF6) and Tax and suppresses interleukin 1β (IL-1β) and Tax-induced NF-κB activation. Recent evidence has shown that USP20 overexpression may impede the proliferation of human T cell leukaemia virus type 1/adult T cell leukaemia (HTLV-1/ATL) cells and suggest that screening for small-molecule compounds to enhance USP20 deubiquitylase activity may unveil new agents that are useful for treating adult T cell leukaemia (Yasunaga et al., 2011).


References

Komander D, Clague MJ and Urbe S (2009) Breaking the chains: structure and function of the deubiquitinases. Nat Rev Mol Cell Biol, 10, 550-563.

Li Z, Wang D, Messing EM and Wu G (2005) VHL protein-interacting deubiquitinating enzyme 2 deubiquitinates and stabilizes HIF-1alpha. EMBO reports, 6, 373-378.

Nagase T, Ishikawa K, Suyama M, Kikuno R, Hirosawa M, Miyajima N, et al. (1999) Prediction of the coding sequences of unidentified human genes. XIII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA research : an international journal for rapid publication of reports on genes and genomes, 6, 63-70.

Reyes-Turcu FE, Ventii KH and Wilkinson KD (2009) Regulation and cellular roles of ubiquitin-specific deubiquitinating enzymes. Annual review of biochemistry, 78, 363-397.

Yasunaga J, Lin FC, Lu X and Jeang KT (2011) Ubiquitin-specific peptidase 20 targets TRAF6 and human T cell leukemia virus type 1 tax to negatively regulate NF-kappaB signaling. Journal of virology, 85, 6212-6219.