title: Probing the Small-Molecule Inhibition of an Anticancer Therapeutic Protein-Protein Interaction Using a Solid-State Nanopore.
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.
authors.affiliation: Structural Biology & Nanopore Research Laboratory, Disease Target Structure Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.|Department of Bio-Analytical Sciences, Korea University of Science and Technology, 217 Gajeong-ro, Yuseong-gu, Daejeon, 34113, Republic of Korea.
authors.forename: Dong-Kyu
authors.initials: DK
authors.lastname: Kwak
authors.affiliation: Department of Materials Science and Engineering, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea.
authors.forename: Hongsik
authors.initials: H
authors.lastname: Chae
authors.affiliation: Structural Biology & Nanopore Research Laboratory, Disease Target Structure Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
authors.forename: Mi-Kyung
authors.initials: MK
authors.lastname: Lee
authors.affiliation: Structural Biology & Nanopore Research Laboratory, Disease Target Structure Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
authors.forename: Ji-Hyang
authors.initials: JH
authors.lastname: Ha
authors.affiliation: School of Biomedical Engineering, Science and Health Systems Drexel University, 3701 Market Street, Philadelphia, PA, 19104, USA.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Goyal
authors.affiliation: School of Biomedical Engineering, Science and Health Systems Drexel University, 3701 Market Street, Philadelphia, PA, 19104, USA.|Department of Mechanical Engineering and Mechanics, Drexel University, 3701 Market Street, Philadelphia, PA, 19104, USA.
authors.forename: Min Jun
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: Department of Materials Science and Engineering, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea. kibum@snu.ac.kr.
authors.forename: Ki-Bum
authors.initials: KB
authors.lastname: Kim
authors.affiliation: Structural Biology & Nanopore Research Laboratory, Disease Target Structure Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea. swchi@kribb.re.kr.|Department of Bio-Analytical Sciences, Korea University of Science and Technology, 217 Gajeong-ro, Yuseong-gu, Daejeon, 34113, Republic of Korea. swchi@kribb.re.kr.
authors.forename: Seung-Wook
authors.initials: SW
authors.lastname: Chi
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000378:metabolism; D001713:Biphenyl Compounds / Q000737:chemistry / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D058608:Nanopores; D009596:Nitrophenols / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010879:Piperazines / Q000737:chemistry / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013449:Sulfonamides / Q000737:chemistry / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C501332:ABT-737; D000970:Antineoplastic Agents; D001713:Biphenyl Compounds; D007093:Imidazoles; D009596:Nitrophenols; D010879:Piperazines; D013449:Sulfonamides; D016159:Tumor Suppressor Protein p53; C482205:nutlin 3; D051736:Proto-Oncogene Proteins c-mdm2; D005982:Glutathione Transferase
keywords: MDM2; drug screening; p53; protein-protein interactions; solid-state nanopores