Product Details
| MW: | ~16kDa |
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| Source: | Produced in E. coli. |
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| UniProt ID: | P61077 |
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| Formulation: | Liquid. In 20 mM Tris-HCl, pH 7.5, containing 0.5 mM DTT. |
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| Purity: | ≥90% (SDS-PAGE) |
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| Biological Activity: | The His6-tagged fusion proteins of UbcH5a, UbcH5b and UbcH5c all charge and support ubiquitinylation in vitro. Unlike their GST-tagged counterparts, the His6-tagged UbcH5 family members all appear to form thiol ester conjugates with ubiquitin at a similar rate under similar conditions. |
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| Application Notes: | Useful for in vitro ubiquitinylation reactions. Typical enzyme concentration to support conjugation in vitro is 100nM to 1μM depending upon conditions. The His-tagged version of this enzyme is not susceptible to self-ubiquitinylation, which can occur with GST-tagged versions. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Scientific Background: | A number of E2s in Saccharomyces cerevisiae and their homologues have been identified. One such family of E2s is the UBC4/5, characterised as essential for the degradation of short-lived, regulatory and abnormal proteins. Protein levels of S. cerevisiae UbC4/5 are up-regulated in response to stress, and their loss results in severe effects on cellular functions.
A human gene product that is 79% identical to S. cerevisiae UBC4/5 in amino-acid sequence was identified as UbcH5a. In addition, two other human members of this highly conserved E2 class were also cloned and designated as UbcH5b and UbcH5c, having 88% and 89% identity to UbcH5a, respectively. Members of the UbcH5a/b/c are the most active class of E2s in cell extracts. The importance of this enzyme class is underscored by the critical role of UBC4/5 in S. cerevisiae. UbcH5a stimulates the conjugation of ubiquitin to the tumour-repressor p53 in the presence of E6-AP and E6. Moreover, UbcH5 family is implicated in c-fos recognition, the modulation of which is controlled by the ubiquitin-proteasome pathway. UbcH5b and UbcH5c are associated with the signal-induced conjugation and subsequent degradation of IkBα in the presence of the SCF complexes. UbcH5c also catalyses the ubiquitination leading to the processing of p105 precursor to form p50, a subunit of the heterodimeric transcription factor NF-kB. The range and diversity of substrates and E3s with which this class of E2 enzymes interact, suggest their complex roles in cellular functions require to be studied further. |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Cullin 4b-RING ubiquitin ligase targets IRGM1 to regulate Wnt signaling and intestinal homeostasis: Y. Fan, et al.; Cell Death Differ. (2022),
Abstract;
CD95/Fas suppresses NF-kB activation through recruitment of KPC2 in a CD95L/FasL-independent mechanism: J.P. Guégan, et al.; iScience
24, 103538 (2021),
Abstract;
Biophysical and biological evaluation of optimized stapled peptide inhibitors of the linear ubiquitin chain assembly complex (LUBAC): F. Aguilar-Alonso, et al.; Bioorg. Med. Chem.
26, 1179 (2018),
Abstract;
BLM Potentiates c-Jun Degradation and Alters Its Function as an Oncogenic Transcription Factor: R. Priyadarshini, et al.; Cell Rep.
24, 947 (2018),
Abstract;
Prp19/Pso4 Is an Autoinhibited Ubiquitin Ligase Activated by Stepwise Assembly of Three Splicing Factors: T.R. de Moura, et al.; Mol. Cell
69, 979 (2018),
Abstract;
ZNRF1 promotes Wallerian degeneration by degrading AKT to induce GSK3B-dependent CRMP2 phosphorylation: S. Wakatsuki, et al.; Nat. Cell Biol.
13, 1415 (2011),
Abstract;
General Literature References
Identification of the ubiquitin carrier proteins, E2s, involved in signal-induced conjugation and subsequent degradation of IkappaBalpha: H. Gonen et al.; J. Biol. Chem. 274, 14823 (1999),
Enzymes catalyzing ubiquitination and proteolytic processing of the p105 precursor of nuclear factor kappaB1. : Coux, O. and Goldberg, A.L.; J. Biol. Chem. 273, 8820-8 (1998),
Degradation of the proto-oncogene c-Fos by the ubiquitin proteolytic system in vivo and in vitro: identification and characterisation of the conjugating enzymes: I. Stancovski et al.; Mol. Cel. Biol. 15, 7106 (1995),
Identification of a family of closely related human ubiquitin conjugating enzymes: J.P. Jensen et al.; J. Biol. Chem. 270, 30408 (1995),
Identification of a human ubiquitin-conjugating enzyme that mediates the E6-AP-dependent ubiquitination of p53. : Scheffner, M.; Proc. Natl. Acad. Sci. USA 91, 8797-8801 (1994),
The ubiquitin-conjugation system. : Jentsch, S. ; Annu. Rev. Genet. 22, 179-207 (1992),
Ubiquitin-conjugating enzymes UBC4 and UBC5 mediate selective degradation of short-lived and abnormal proteins. : Seufert, W. and Jentsch, S.; EMBO J. 9, 543-50 (1990),
