Recombinant Human His6-USP8 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human His6-USP8 Protein, CF Summary

• Details of Functionality: Recombinant Human His6-USP8 is a Ubiquitin-specific deconjugating enzyme. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human His6-USP8 concentration of 10-50 nM.
• Source: Spodoptera frugiperda, Sf 9 (baculovirus)-derived human USP8 protein
  Met1 - Thr1118 (Pro2Gly, Val4Ala & V257Ala) with a N-terminal 6-His tag
• Accession #: P40818.1
• Protein/Peptide Type: Recombinant Enzymes
• Gene: USP8
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 131 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
• Buffer: Supplied as a solution in HEPES, NaCl and TCEP.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human His6-USP8 Protein, CF

• Deubiquitinating enzyme 8
• EC 3.1.2.15
• EC 3.4.19.12
• HumORF8
• KIAA0055hUBPy
• MGC129718
• ubiquitin carboxyl-terminal hydrolase 8
• Ubiquitin isopeptidase Y
• ubiquitin specific peptidase 8
• ubiquitin specific protease 8
• ubiquitin thioesterase 8
• Ubiquitin thiolesterase 8
• Ubiquitin-specific-processing protease 8
• UBPY
• UBPYFLJ34456
• USP8

Background

Ubiquitin Specific Peptidase 8 (USP8), also known as Ubiquitin Isopeptidase Y (UBPY), is a widely expressed deubiquitinating enzyme belonging to the peptidase C19 family. It has a predicted molecular weight of 127.5 kDa (1). Human USP8 is 1118 amino acids (aa) in length and shares 84% aa sequence identity with the mouse and rat orthologs (2). It contains an N-terminal MIT domain (aa 33-116) that mediates endosomal localization, CHMP-binding, and maintenance of ESCRT-0 (3). USP8 also has a rhodanese domain (aa 181-319) that binds NRDP1 Ubiquitin ligase (E3), a SH3 domain binding sequence (aa 405-413), and a C-terminal catalytic domain (aa 734-1110) (2,4). USP8 is a growth-regulated enzyme that controls the internalization and endocytic trafficking of cell surface receptors (1,5). Some receptors are targeted for internalization and degradation by ubiquitination. USP8 has been shown to disrupt the down-regulation of multiple receptors, including EGF R/ErbB1, ErbB2/Her2, and Smoothened, via their deubiquitination (6-9). Conversely, USP8 appears to have the opposite effect on the trafficking of CXCR4, PAR2, and the δ-opioid receptor (10-12). Depletion or catalytic inactivation of USP8 stabilized their expression (10-12). It is thought that deubiquitination of these receptors down-stream of the sorting endosome commits them to lysosomal degradation (10). USP8 can be phosphorylated at Ser680 allowing for 14-3-3-epsilon binding, which subsequently inhibits USP8 activity (13). Additionally, USP8 undergoes tyrosine phosphorylation at its N-terminus following EGF activation of the EGF R/ErbB1 / ErbB2/Her2 receptor complex (7). 

1. Naviglio, S et al. (1998) EMBO J. 17:3241.
2. Avvakumov, G.V. et al. (2006) J. Biol. Chem. 281:38061.
3. Row, P.E. et al. (2007) J. Biol. Chem. 282:30929.
4. Kato, M. et al. (2000) J. Biol. Chem. 275:37481.
5. Wright, M.H. et al. (2011) Cell Biochem. Biophys. 60:39.
6. Mizuno, E. et al. (2005) Mol. Biol. Cell 16:5163.
7. Meijer, I. M. and J.E. van Leeuwen (2011) Cell. Signal. 23:458.
8. Xia, R. et al. (2012) PLoS Biol. 10:e1001238.
9. Li, E. et al. (2012) PLoS Biol. 10:e1001239.
10. Hislop, J.N. et al. (2009) J. Biol. Chem. 284:19361.
11. Hasdemir, B. et al. (2009) J. Biol. Chem. 284:28453.
12. Berlin, I. et al. (2010) J. Biol. Chem. 285:37895.
13. Mizuno, E. et al. (2007) Exp. Cell Res. 313:3624.

Publications for UBPY/USP8 (E-520)(3)

Publications using E-520

• Jadhav, P;Liang, X;Ansari, A;Tan, B;Tan, H;Li, K;Chi, X;Ford, A;Ruiz, FX;Arnold, E;Deng, X;Wang, J; Design of quinoline SARS-CoV-2 papain-like protease inhibitors as oral antiviral drug candidates Nature communications 2025-02-13 [PMID: 39948104] (Bioassay, N/A)
• Papp , Stephani, Huber , Anne-Lau, Jordan , Sabine D, Kriebs , Anna, Nguyen , Madelena, Moresco , James J, Yates , John R, Lamia , Katja A DNA damage shifts circadian clock time via Hausp-dependent Cry1 stabilization. Elife, 2015-03-10;4(0):. 2015-03-10 [PMID: 25756610] (Bioassay, Human)
• Liang , Qin, Dexheimer , Thomas S, Zhang , Ping, Rosenthal , Andrew S, Villamil , Mark A, You , Changjun, Zhang , Qiuting, Chen , Junjun, Ott , Christin, Sun , Hongmao, Luci , Diane K, Yuan , Bifeng, Simeonov , Anton, Jadhav , Ajit, Xiao , Hui, Wang , Yinsheng, Maloney , David J, Zhuang , Zhihao A selective USP1-UAF1 inhibitor links deubiquitination to DNA damage responses. Nat Chem Biol, 2014-02-16;10(4):298-304. 2014-02-16 [PMID: 24531842] (Bioassay)

Bioinformatics

• Gene Symbol: USP8
• Uniprot:
  • P40818.1()