Recombinant Human His6-MBP-HOIP/RNF31 Catalytic Domain, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human His6-MBP-HOIP/RNF31 Catalytic Domain, CF Summary

• Details of Functionality:

  Recombinant Human HOIP/RNF31 is a RING domain Ubiquitin ligase (E3) and component of the LUBAC complex that functions downstream of a Ubiquitin-activating (E1) enzyme and a Ubiquitin-conjugating (E2) enzyme to conjugate Ubiquitin to substrate proteins. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HOIP/RNF31 concentration of 0.1-0.5 μM.

• Source: E. coli-derived human HOIP/RNF31 protein
  Contains an N-terminal 6-His and MBP (Maltose Binding Protein) tag
• Accession #: Q96EP0
• Protein/Peptide Type: Recombinant Enzymes
• Gene: RNF31
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain

Applications/Dilutions

• Theoretical MW: 87 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.
  • 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 7.5, 150 mM NaCl, 10% Glycerol, 1 mM TCEP

• Purity: >90%, 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-MBP-HOIP/RNF31 Catalytic Domain, CF

• EC 6.3.2.-
• FLJ10111
• FLJ23501
• HOIP
• MGC19975
• ring finger protein 31
• RNF31
• ZIBRA
• ZIBRAHOIL-1-interacting protein
• Zinc in-between-RING-finger ubiquitin-associated domain protein

Background

HOIP (HOIL-1 Interacting Protein, also known as RNF31) is a RING domain ubiquitin E3 ligase and component of the LUBAC complex. LUBAC, a large (approximately 600 kDa) heteromultimeric assembly consisting of at least HOIP, HOIL-1, and Sharpin, conjugates linear polyubiquitin chains to various substrates and plays a key role in NF-kappa B activation and signaling events mediated through TNF alpha and IFN. Both the catalytic activity and specificity for linear ubiquitin chain formation of LUBAC are contained within the C-terminal RBR (RING-IBR-RING) and LDD (linear ubiquitin chain determining domain) domains of HOIP. HOIL-1 and Sharpin are essential for full-length HOIP activity, but not for HOIP truncation mutants consisting of the RBR and LDD domains. This protein contains N-terminal 6-His and MBP tags, and is useful for generating unanchored linear polyubiquitin chains without the requirement for HOIL-1 or Sharpin. HOIP catalytic domain contains aa’s 699-1072.

1. Belgnaoui S.M., et al. (2012) Cell Host Microbe 12: 211-222
2. Smit J.J, et al. (2012) EMBO J. 31: 3833-3844
3. Stieglitz B., et al. (2012) EMBO Rep. 13: 840-846
4. Wauer T. and Komander D. (2013) EMBO J. doi:10.1038/emboj.2013.125

Bioinformatics

• Gene Symbol: RNF31
• Uniprot:
  • Q96EP0()