Probing the Small‐Molecule Inhibition of an Anticancer Therapeutic Protein‐Protein Interaction Using a Solid‐State Nanopore |
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| Authors: | Dong‐Kyu Kwak Hongsik Chae Dr Mi‐Kyung Lee Dr Ji‐Hyang Ha Dr Gaurav Goyal Prof Min Jun Kim Prof Ki‐Bum Kim Dr Seung‐Wook Chi |
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| Institution: | 1. Structural Biology & Nanopore Research Laboratory, Disease Target Structure Research Center, KRIBB, Daejeon, Republic of Korea;2. Department of Bio-Analytical Sciences, Korea University of Science and Technology, Daejeon, Republic of Korea;3. Department of Materials Science and Engineering, Seoul National University, Seoul, Republic of Korea;4. School of Biomedical Engineering, Science and Health Systems Drexel University, Philadelphia, PA, USA;5. Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, PA, USA |
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| Abstract: | 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. |
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| Keywords: | drug screening MDM2 p53 protein– protein interactions solid-state nanopores |
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