title: Enzyme-catalyzed assembly of gold nanoparticles for visualized screening of DNA base excision repair.
abstract: Activity screening of DNA base excision repair (BER) enzymes is a crucial step for understanding numerous fundamental biochemical processes. A novel label-free homogeneous technique is developed for visualized uracil-DNA glycosylase (UDG) activity assay using gold nanoparticles (AuNPs). This strategy relies on the enzyme-catalyzed assembly of AuNPs decorated with DNA probes. In the presence of endonuclease IV (an enzyme which can further hydrolyze the products from UDG-catalyzed reaction), the substrate DNA selectively interacts with UDG followed by the efficient release of a single-strand probe. The released single-strand probe then makes the network-like assembly of decorated AuNPs to provide a visible signal for UDG activity. This strategy that can be performed in a label-free homogeneous assay format improved the duration, the simplicity and the throughput of UDG activity screening. The results provided in the present study revealed that this strategy could hold great potential as a robust, convenient and visualized platform for activity screening of uracil-DNA glycosylases with high selectivity and desirable sensitivity.
authors.affiliation: State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Van-Trong
authors.initials: VT
authors.lastname: Nguyen
authors.forename: Dinh-Vu
authors.initials: DV
authors.lastname: Le
authors.forename: Chan
authors.initials: C
authors.lastname: Nie
authors.forename: Dian-Ming
authors.initials: DM
authors.lastname: Zhou
authors.forename: Yu-Zhi
authors.initials: YZ
authors.lastname: Wang
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Tang
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
authors.forename: Ru-Qin
authors.initials: RQ
authors.lastname: Yu
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004260:DNA Repair; D057075:Enzyme Assays / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D051981:Uracil-DNA Glycosidase / Q000378:metabolism*
chemical_list: D015342:DNA Probes; D006046:Gold; D004247:DNA; D051981:Uracil-DNA Glycosidase