USP4 [6His-tagged]


Catalogue Number
64-0001-050
Product Size
50 µg
Price £
£325
Accession Number
NP_003354
Residues Expressed
1-963
Certificate of Analysis Size
50 µg
Species
human
Source
E.coli
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
~111 kDa
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: NP_003354. 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 His-USP4 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 His-USP4 was compared confirming the deubiquitylating activity of His-USP4.


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 carboxyl-terminal hydrolase 4 (Ubiquitin Specific Protease 4; USP4) is a member of the cysteine protease enzyme family and cloning of the human gene was first described by Gupta et al. (1993). In 1995, USP4 was identified as a proto-oncogene related to USP6, showing a consistently elevated gene expression level in small cell tumours and lung adenocarcinomas suggesting that it may have a possible causative role in neoplasia (Gray et al., 1995). USP4 has been implicated in a number of other processes, including protein quality control in the endoplasmatic reticulum and p53 and Wnt signalling. USP4 has also been reported to inhibit the kinase TAK1 that is ubiquitylated by the AKT regulator TRAF6 (Uras et al., 2012).


References

Gray DA, Inazawa J, Gupta K, Wong A, Ueda R and Takahashi T (1995) Elevated expression of Unph, a proto-oncogene at 3p21.3, in human lung tumors. Oncogene 10, 2179-2183.

Gupta K, Copeland NG, Gilbert DJ, Jenkins NA and Gray DA (1993) Unp, a mouse gene related to the tre oncogene. Oncogene 8, 2307-2310.

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.

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

Uras IZ, List T and Nijman SM (2012) Ubiquitin-specific protease 4 inhibits mono-ubiquitination of the master growth factor signalling kinase PDK1. PloS one 7,e31003.