Product Details
| Alternative Name: | Human double minute 2 protein |
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| MW: | ~35kDa |
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| Source: | Produced in E. coli BL21 (λDE3). |
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| UniProt ID: | Q00987 |
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| Formulation: | Liquid. In TBS, pH 7.5, containing 150mM sodium chloride and 1mM DTT. |
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| Purity: | ≥95% (SDS-PAGE) |
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| Purity Detail: | Purified by glutathione affinity chromatography. |
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| Activity: | Modifies p53 (Prod. No. BML-FW8820) in ubiquitinylation reactions containing Ube1 (Prod. No. BML-UW9410), UbcH5b (Prod. No. BML-UW9060) and energy regeneration solution. |
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| Application Notes: | Uses:
1.in vitro generation of ubiquitinylated p53 for use in Ub-p53 related studies.
2.Investigation of the p53 ubiquitin modification process itself.
3.Ubiquitin modification of other known Hdm2 substrate proteins.
4.NEDD8 modification of p53. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Scientific Background: | p53 is a much studied and complex multifunctional protein, which plays a major role in the cellular response to DNA damage and other genomic aberrations. The activation of p53 can lead to either cell cycle arrest and DNA repair, or apoptosis, through its involvement in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for these processes. Activation and regulation of the p53 transcription pathway is controlled by a range of post-translational modifications including acetylation, phosphorylation and ubiquitinylation. In normal cells, p53 is maintained at a low level mainly through Hdm2-mediated ubiquitinylation and subsequent degradation by the proteasome. Hdm2 is a RING domain dependent ubiquitin E3 ligase that utilizes its C-terminal RING domain to promote not only p53 ubiquitinylation, predominantly at the C-terminus of p53, but also to target Hdm2 itself for auto-ubiquitinylation and subsequent degradation. The isolated Hdm2 C-terminal RING domain (residues 418-491) has been shown to be sufficient for both p53 and self-ubiquitinylation activity. |
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| Regulatory Status: | RUO - Research Use Only |
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Activity analysis: Hdm2 catalytic RING domain ubiquitinylation of p53 (Prod. No. BML-FW8820) +/- Hdm2 RING domain. p53 species detected by western blotting using p53 monoclonal antibody (Prod. No. BML-PW1085).
SDS-PAGE analysis: Lane 1: 2μg GST-Hdm2 catalytic RING domain (Prod. No. BML-UW0200). Lane 2: MW markers (top to bottom) 116, 97, 84, 66, 55, 45, 36, 29, 24, 20.
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Product Literature References
The ubiquitin ligase LIN41/TRIM71 targets p53 to antagonize cell death and differentiation pathways during stem cell differentiation: D.T.T. Nguyen, et al.; Cell Death Differ.
24, 1063 (2017),
Abstract;
Full Text
General Literature References
Solution Structure of the Hdm2 C2H2C4 RING, a Domain Critical for Ubiquitination of p53: M. Kostic, et al.; J. Mol. Biol. (2006),
Ubiquitination, phosphorylation and acetylation: the molecular basis for p53 regulation: C.L. Brooks, et al.; Curr. Opin. Cell Biol. 15, 164 (2003),
Deconstruction of p53 functions and regulation: Y. Haupt, et al.; Oncogene 21, 8223 (2002),
Mdm2 is a RING finger-dependent ubiquitin protein ligase for itself and p53: S. Fang, et al.; J. Biol. Chem. 275, 8945 (2000),
Multiple lysine mutations in the C-terminal domain of p53 interfere with MDM2-dependent protein degradation and ubiquitination: S. Nakamura, et al.; Mol. Cell Biol. 20, 9391 (2000),
Regulation of the p53 tumor suppressor protein: M. Oren, et al.; J. Biol. Chem. 274, 36031 (1999),
Mdm2 promotes the rapid degradation of p53: Y. Haupt, et al.; Nature 387, 296 (1997),
Isolation and characterization of a human p53 cDNA clone: expression of the human p53 gene: G. Matlashewski, et al.; EMBO J. 3, 3257 (1984),
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