A Tailored Phospho‐p53 Library Probes Antibody Specificity and Recognition Limitations
Mateusz Hess, Jonathan H. Davies, Sofia Margiola, Sonja Schneider, Thomas Hicks, Nishant Rai, Manuel M. Müller
TL;DR
Researchers created a library of phosphorylated p53 proteins to test antibody specificity and found that phosphorylation can hide antibody binding sites.
Contribution
A tailored library of site-specifically phosphorylated p53 proteins was developed to probe antibody recognition and epitope masking.
Findings
The study validated the specificity of phosphorylation-specific anti-p53 antibodies.
Phosphorylation was found to cause widespread epitope masking in commonly used antibodies.
Abstract
The tumor suppressor protein p53, known as the “guardian of the genome,” is regulated by a complex network of post‐translational modifications. Phosphorylations at 7 Ser/Thr residues within the N‐terminal transactivation domain 1 (TAD1) play a role in p53 activation, yet their precise mechanisms of action remain elusive due to challenges in accessing well‐defined phosphorylated isoforms. To address this limitation, this study harnesses a recently developed approach for the semisynthesis of site‐specifically phosphorylated p53 to generate a comprehensive library of singly phosphorylated p53 including all TAD1 sites: Ser6, Ser9, Ser15, Thr18, Ser20, Ser33, and Ser37. The library was then used to probe the specificity of common p53 antibodies in western blot analysis. This study's results confirm the specificity of the target site of most phosphorylation‐specific anti‐p53 antibodies, but…
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Taxonomy
TopicsChemical Synthesis and Analysis · Advanced biosensing and bioanalysis techniques · Cancer-related Molecular Pathways