Probing the Small-Molecule Inhibition of an Anticancer Therapeutic Protein-Protein Interaction Using a Solid-State Nanopore

• Published in: Angewandte Chemie Vol. 128; no. 19; pp. 5807 - 5811
• Main Authors: Kwak, Dong-Kyu, Chae, Hongsik, Lee, Mi-Kyung, Ha, Ji-Hyang, Goyal, Gaurav, Kim, Min Jun, Kim, Ki-Bum, Chi, Seung-Wook
• Format: Journal Article
• Language: English
• Published: Weinheim Blackwell Publishing Ltd 04.05.2016; Wiley Subscription Services, Inc
• Subjects: Blocking; Cancer; Charging; Chemistry; Conversion; Electric potential; Festkörper-Nanoporen; Inhibition; MDM2; Nanostructure; p53; Platforms; Protein-Protein-Wechselwirkungen; Proteins; Voltage; Wirkstoff-Screening
• ISSN: 0044-8249; 1521-3757
• DOI: 10.1002/ange.201511601

Nanopore sensing is an emerging technology for the single‐molecule‐based detection of various biomolecules. In this study, we probed the anticancer therapeutic p53 transactivation domain (p53TAD)/MDM2 interaction and its inhibition with a small‐molecule MDM2 antagonist, Nutlin‐3, using low‐noise solid‐state nanopores. Although the translocation of positively charged MDM2 through a nanopore was detected at the applied negative voltage, this MDM2 translocation was almost completely blocked upon formation of the MDM2/GST‐p53TAD complex owing to charge conversion. In combination with NMR data, the nanopore measurements showed that the addition of Nutlin‐3 rescued MDM2 translocation, indicating that Nutlin‐3 disrupted the MDM2/GST‐p53TAD complex, thereby releasing MDM2. Taken together, our results reveal that solid‐state nanopores can be a valuable platform for the ultrasensitive, picomole‐scale screening of small‐molecule drugs against protein–protein interaction (PPI) targets. Die Protein‐Protein‐Wechselwirkung zwischen MDM2 und der p53‐Transkriptionsaktivierungsdomäne (GST‐p53TAD) sowie die Hemmung dieser Wechselwirkung durch Nutlin‐3 wurden mithilfe einer rauscharmen Festkörper‐Nanopore detektiert. Dieses Nanoporensystem könnte zum Hochdurchsatz‐Screening anderer Inhibitoren von Protein‐Protein‐Wechselwirkungen genutzt werden.