His6-UBE2K, human recombinant

Catalog number: SBB-CE0022, 100 μg

Recombinant Human UBE2K (HIP2). E2 enzyme which catalyzes free K48-linked polyubiquitin chains. Known to interact with huntingtin and TRIM6. The protein is N-terminally His-tagged, and recombinantly expressed in E.Coli. [1][2]

In stock
SKU
0022
$179.00

Description

Human UBE2K (HIP-2) is an E2 ubiquitin conjugating enzyme. An E1 activating enzyme is required to attach ubiquitin to UBE2K via an active site cysteine. The mechanism of ubiquitin transfer involves the breaking of a E1-Ub thioester linkage, followed by a reformation of a UBE2K-Ub thioester. UBE2K is capable of synthesizing K-48 linked ubiquitin chains and can do so without an E3 ubiquitin ligating enzyme present. Although an E3 ligase is not required for chain formation, free UBE2K synthesized K48 polyubiquitin chains have been shown to interact with the E3 TRIM6, leading to activation of IKKe kinase activity and subsequent antiviral activity. This recombinant UBE2K is expressed in E.coli with an N-terminal poly-histidine tag. Final purity for this enzyme is > 95% - determined by SDS-PAGE.

For Research Use Only, Not For Use In Humans.

Specifications

Product Overview
Quantity: 100 μg
Molecular Weight: 22 kDa
Purity: >95% by SDS-PAGE
Substrate Properties: Working concentrations of this enzyme range from 1 to 5µM
Storage Buffer: HEPES pH 7.5, 150mM NaCl, 10% glycerol, 2mM TCEP
Solubility Store at −80°C after product arrival. Avoid multiple freeze / thaws. It is recommended to make multiple aliquots after the first thaw.

Figures & Data

UBE2K, human recombinant

Figure 1. UBE2K, SDS-PAGE From left to right, increasing amounts of UBE2K loaded onto a 4-20% SDS-PAGE gel, stained with Coomassie Brilliant Blue. UBE2K is > 95% pure.

E2 Thioester Activity

Figure 2. Thioester Activity Assay. UBE2K forms a thioester with UB in an ATP dependent manner, and the bond can be reduced with addition of excess DTT. The thioester assay also shows di-ubiquitin formation with addition of ATP. The UBE2K is active.

Citations & References

1) Komander, David and Michael Rape. "The Ubiquitin Code". Annual Review of Biochemistry 81.1 (2012): 203-229. Web. 9 Mar. 2017.

2) Middleton, Adam J. and Catherine L. Day. "The Molecular Basis Of Lysine 48 Ubiquitin Chain Synthesis By Ube2k". Scientific Reports 5 (2015): 16793. Web. 9 Mar. 2017.