title: Nanosensing protein allostery using a bivalent mouse double minute two (MDM2) assay.
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.
authors.affiliation: Centre for Molecular Nanometrology, WestCHEM, Department of Pure and Applied Chemistry,  University of Strathclyde, Glasgow G1 1XL,  United Kingdom.
authors.forename: Anna F
authors.initials: AF
authors.lastname: Robson
authors.forename: Ted R
authors.initials: TR
authors.lastname: Hupp
authors.forename: Fiona
authors.initials: F
authors.lastname: Lickiss
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Ball
authors.forename: Karen
authors.initials: K
authors.lastname: Faulds
authors.forename: Duncan
authors.initials: D
authors.lastname: Graham
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D019281:Dimerization; D015195:Drug Design; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D036103:Nanotechnology / Q000379:methods*; D009369:Neoplasms / Q000175:diagnosis / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D054730:Protein Interaction Domains and Motifs / Q000502:physiology; D012834:Silver / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013059:Spectrum Analysis, Raman / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C415788:Mtbp protein, mouse; D016159:Tumor Suppressor Protein p53; D012834:Silver