Nanosensing protein allostery using a bivalent mouse double minute two (MDM2) assay

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Robson, Anna F. and Hupp, Ted R. and Lickiss, Fiona and Ball, Kathryn L. and Faulds, Karen and Graham, Duncan (2012) Nanosensing protein allostery using a bivalent mouse double minute two (MDM2) assay. Proceedings of the National Academy of Sciences, 109 (21). pp. 8073-8078. ISSN 0027-8424 (https://doi.org/10.1073/pnas.1116637109)

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Abstract

The tumor suppressor protein, p53, is either mutated or absent in >50% of cancers and is negatively regulated by the mouse double minute (MDM2) protein. Understanding and inhibition of the MDM2-p53 interaction are, therefore, critical for developing novel chemotherapeutics, which are currently limited because of a lack of appropriate study tools. We present a nanosensing approach to investigate full-length MDM2 interactions with p53, thus providing an allosteric assay for identifying binding ligands. Surface-enhanced Raman scattering (SERS)-active nanoparticles, functionalized with a p53 peptide mimic (peptide 12.1), display biologically specific aggregation following addition of MDM2. Nanoparticle assembly is competitively inhibited by the N-terminal MDM2-binding ligands peptide 12.1 and Nutlin-3. This study reports nanoparticle assembly through specific protein-peptide interactions that can be followed by SERS. We demonstrate solution-based MDM2 allosteric interaction studies that use the full-length protein.

ORCID iDs

Robson, Anna F., Hupp, Ted R., Lickiss, Fiona, Ball, Kathryn L., Faulds, Karen ORCID logoORCID: https://orcid.org/0000-0002-5567-7399 and Graham, Duncan ORCID logoORCID: https://orcid.org/0000-0002-6079-2105;
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