title: Probing the Dynamic Interaction between Damaged DNA and a Cellular Responsive Protein Using a Piezoelectric Mass Biosensor.
abstract: The binding events between damaged DNA and recognition biomolecules are of great interest for understanding the activity of DNA-damaging drugs and the related DNA repair networks. Herein, a simple and sensitive sensor system was tailored for real-time probing of the dynamic molecular recognition between cisplatin-damaged-DNA (cisPt-DNA) and a cellular responsive protein, high-mobility-group box 1 (HMGB1). By integration of flow injection analysis (FIA) with quartz crystal microbalance (QCM), the interaction time-course of cisPt-DNA and HMGB1 domain A (HMGB1a) was investigated. The highly specific sensing interface was carefully designed and fabricated using cisPt-DNA as recognition element. A hybrid self-assembled monolayer consisting of cysteamine and mercaptohexanol was introduced to resist nonspecific adsorption. The calculated kinetic parameters (kassand kdiss) and the dissociation constant (KD) demonstrated the rapid recognition and tight binding of HMGB1a toward cisPt-DNA. Molecular docking was employed to simulate the complex formed by cisPt-DNA and HMGB1a. The tight binding of such a DNA-damage responsive complex is appealing for the downstream molecular recognition event related to the resistance to DNA repair. This continuous-flow QCM biosensor is an ideal tool for studying specific interactions between drug-damaged-DNAs and their recognition proteins in a physiological-relevant environment, and will provide a potential sensor platform for rapid screening and evaluating metal anticancer drugs.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Kui
authors.identifier: http://orcid.org/0000-0002-4995-0395
authors.initials: K
authors.lastname: Wu
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Fuyi
authors.identifier: http://orcid.org/0000-0003-0962-1260
authors.initials: F
authors.lastname: Wang
authors.affiliation: Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Rui
authors.identifier: http://orcid.org/0000-0001-7191-9354
authors.initials: R
authors.lastname: Zhao
mesh_terms: D015374:Biosensing Techniques; D004247:DNA; D062105:Molecular Docking Simulation; D011506:Proteins; D011791:Quartz; D058525:Quartz Crystal Microbalance Techniques
chemical_list: D011506:Proteins; D011791:Quartz; D004247:DNA
keywords: DNA damage; high-mobility-group box 1; kinetic analysis; molecular recognition; piezoelectric mass biosensor