Recombinant Human Isopeptidase T/USP5 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Isopeptidase T/USP5 Protein, CF Summary

Details of Functionality
Recombinant Human Isopeptidase T/USP5 is a Ubiquitin-specific deconjugating enzyme. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Isopeptidase T/USP5 concentration of 10-100 nM.  A 15 minute pre-incubation with 10 mM DTT is recommended to achieve maximum activity.
Source
E. coli-derived human Isopeptidase T/USP5 protein
Accession #
Protein/Peptide Type
Recombinant Enzymes
Gene
USP5
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain

Applications/Dilutions

Theoretical MW
96 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.
Publications
Read Publications using
E-322 in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
X mg/ml (X μM) in 50 mM HEPES pH 8.0, 150 mM NaCl, 1mM DTT
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 Isopeptidase T/USP5 Protein, CF

  • Deubiquitinating enzyme 5
  • EC 3.1.2.15
  • EC 3.4.19.12
  • Isopeptidase T
  • IsoT
  • ISOTubiquitin carboxyl-terminal hydrolase 5
  • ubiquitin isopeptidase T
  • ubiquitin specific peptidase 5 (isopeptidase T)
  • ubiquitin specific protease 5 (isopeptidase T)
  • ubiquitin thioesterase 5
  • Ubiquitin thiolesterase 5
  • ubiquitin-specific protease-5 (ubiquitin isopeptidase T)
  • Ubiquitin-specific-processing protease 5
  • USP5

Background

Isopeptidase T/Ubiquitin Specific Peptidase 5 (USP5) is a widely expressed deubiquitinating enzyme belonging to the peptidase C19 family (1). It is the only known member that requires zinc binding to be active (2). It has a predicted molecular weight of 95.8 kDa (3). Human Isopeptidase T/USP5 is 858 amino acids (aa) in length and shares 98% aa sequence identity with the mouse and rat orthologs (3). Isopeptidase T/USP5 is largely responsible for the disassembly of unanchored poly-Ubiquitin chains. It binds multiple Ubiquitin molecules in a poly-Ubiquitin chain and can cleave Lys29-, Lys48- and Lys63-linked chains (1,4,5). It contains four putative Ubiquitin-binding domains: an N-terminal zinc finger Ubiquitin-binding (ZnF-UBP) domain, a Ubiquitin-specific processing protease (UBP) catalytic domain, and two Ubiquitin-associated domains (UBA1 and UBA2) (5-7). The ZnF-UBP domain (aa 163-291) selectively interacts with an unmodified C-terminus of poly-Ubiquitin chains and induces a conformational change that prevents Isopeptidase T/USP5 from disassembling poly-Ubiquitin until another deubiquitinating enzyme has released the chain from the ubiquitinated protein (5,6,8). The UBP domain binds the second Ubiquitin in poly-Ubiquitin, while the subsequent Ubiquitins bind the UBA2 (aa 722-762) and UBA1 (aa 654-695) domains (5,7). There are short and long forms of human Isopeptidase T/USP5 that differ by an insertion of 23 aa in the long form (2). Suppression of Isopeptidase T/USP5 has been shown to increase the amount and transcriptional activity of p53 due to the accumulation of unanchored poly-Ubiquitin (9). Conversely, an up-regulation of USP5 has been associated with fetal Down syndrome (10).

  1. Wilkinson, K.D. et al. (1995) Biochemistry 34:14535.
  2. Gabriel, J.M. et al. (2002) Biochemistry 41:13755.
  3. Falquet, L. et al. (1995) FEBS Lett. 376:233.
  4. Raasi, S. et al. (2005) Nat. Struct. Mol. Biol. 12:708.
  5. Reyes-Turcu, F.E. et al. (2009) Annu. Rev. Biochem. 78:363.
  6. Reyes-Turcu, F.E. et al. (2006) Cell 124:1197.
  7. Reyes-Turcu, F.E. et al. (2008) J. Biol. Chem. 283:19581.
  8. Avvakumov, G.V. et al. (2012) Biochemistry 51:1188.
  9. Dayal, S. et al. (2009) J. Biol. Chem. 284:5030.
  10. Engidawork, E. et al. (2001) J. Neural Transm. Suppl.(61):117.

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Publications for Isopeptidase T/USP5 (E-322)(3)

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Bioinformatics

Gene Symbol USP5
Uniprot