P53 Protein (TP53): Target Overview, Research Applications, and Selection Guide
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P53 (tumor protein 53, encoded by the TP53 gene) is a transcription factor and the most frequently mutated tumor suppressor in human cancers. Often called the “guardian of the genome,” p53 integrates multiple stress signals—DNA damage, oncogene activation, hypoxia, and ribosome stress—to induce cell cycle arrest, DNA repair, apoptosis, or senescence. Recombinant p53 proteins are essential tools for studying DNA binding, transcriptional activity, protein-protein interactions (e.g., p53-MDM2, p53-p14ARF), ubiquitination, and small-molecule reactivators of mutant p53. A tp53 recombinant protein (especially p53 protein human and tp53 active protein) is critical for functional assays that require DNA-binding competence or transcriptional activity.
P53 Target Overview
P53 is a tetrameric transcription factor with a well-defined domain structure: an N-terminal transactivation domain (TAD), a proline-rich region, a central DNA-binding domain (DBD), a tetramerization domain (TD), and a C-terminal regulatory domain. Key aspects of p53 biology include:
- Activation by post-translational modifications (phosphorylation, acetylation, ubiquitination) in response to cellular stress
- Regulation of hundreds of target genes involved in cell cycle (p21, 14-3-3σ), apoptosis (PUMA, Bax, Noxa), DNA repair, and metabolism
- Inactivation by MDM2 through direct binding, ubiquitination, and proteasomal degradation
- Frequent mutation in >50% of human cancers, mostly missense mutations in the DNA-binding domain (e.g., R175H, R248W, R273H)
- Gain-of-function (GOF) activities for certain mutants, promoting metastasis and chemoresistance
- Existence of multiple isoforms (Δ40p53, Δ133p53, p53β) with distinct regulatory roles
Because p53 activity is highly context-dependent, selecting the right tp53 recombinant protein (full-length, domain, wild-type vs mutant, active vs unmodified) is critical for experimental success.
P53 Target Information
Target Name: Cellular tumor antigen p53 (tumor protein 53)
Synonyms: TP53, p53, tumor suppressor p53, transformation-related protein 53 (TRP53)
UniProt ID: P04637 (human p53)
Target Class: Transcription factor; tumor suppressor; stress-responsive regulator
Key Research Applications of P53 Protein
Recombinant p53 is widely used in biochemical, biophysical, and cell-based studies. For DNA-binding or transcriptional assays, a tp53 active protein (tetrameric, DNA-binding competent) is required.
DNA-Binding Assays
- Study sequence-specific binding to p53 response elements.
- Support EMSA, fluorescence polarization, SPR, and BLI workflows.
- Evaluate wild-type, mutant, or reactivated p53 proteins.
Protein-Protein Interaction Studies
- Analyze p53-MDM2 binding.
- Study interactions with p14ARF, p300/CBP, 53BP1, and other regulatory proteins.
- Map interaction domains using full-length or domain-specific p53 constructs.
Transcriptional Activity Assays
- Support p53-dependent reporter assay systems.
- Study transcriptional regulation using p53 response elements.
- Evaluate wild-type or mutant p53 transcriptional activity.
Ubiquitination and Degradation Assays
- Use p53 as a substrate for MDM2-mediated ubiquitination.
- Study proteasomal degradation pathways.
- Evaluate E1, E2, ubiquitin, ATP, and MDM2-dependent workflows.
Inhibitor and Reactivator Screening
- Screen compounds that disrupt p53-MDM2 binding.
- Evaluate small molecules that restore DNA-binding activity to mutant p53.
- Support AlphaScreen, TR-FRET, SPR, BLI, and functional DNA-binding assays.
Antibody Development and Validation
- Validate antibodies recognizing wild-type or mutant p53.
- Support detection of p53 post-translational modifications.
- Provide controls for Western blot, IP, IHC, ChIP, or related applications.
P53 Experimental Workflows and Use Cases
Recombinant p53 proteins are most effective in functional and biochemical workflows:
- EMSA – Validate sequence-specific DNA binding of wild-type or reactivated mutant p53
- SPR / BLI – Kinetic analysis of p53-DNA or p53-MDM2 interactions
- AlphaScreen / TR-FRET – High-throughput screening for p53-MDM2 disruptors or p53 reactivators
- In vitro transcription – Measure p53-driven transcription using RNA polymerase II and nuclear extracts
- Ubiquitination assays – Use p53 as a substrate together with MDM2, E1, E2 (UbcH5b) and ubiquitin
- Pull-down assays – Map interaction domains with co-activators (p300) or repressors (MDM2)
Because p53 is a nuclear transcription factor, it is not suitable for membrane-protein or receptor-ligand workflows. For functional DNA binding, ensure you order a tp53 active protein that is verified for tetramerization and sequence-specific DNA binding.
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