title: DNA-conjugated gold nanoparticles based colorimetric assay to assess helicase activity: a novel route to screen potential helicase inhibitors.
abstract: Helicase are essential enzymes which are widespread in all life-forms. Due to their central role in nucleic acid metabolism, they are emerging as important targets for anti-viral, antibacterial and anti-cancer drugs. The development of easy, cheap, fast and robust biochemical assays to measure helicase activity, overcoming the limitations of the current methods, is a pre-requisite for the discovery of helicase inhibitors through high-throughput screenings. We have developed a method which exploits the optical properties of DNA-conjugated gold nanoparticles (AuNP) and meets the required criteria. The method was tested with the catalytic domain of the human RecQ4 helicase and compared with a conventional FRET-based assay. The AuNP-based assay produced similar results but is simpler, more robust and cheaper than FRET. Therefore, our nanotechnology-based platform shows the potential to provide a useful alternative to the existing conventional methods for following helicase activity and to screen small-molecule libraries as potential helicase inhibitors.
authors.affiliation: Nano Innovation Laboratory, Elettra-Sincrotrone Trieste S.C.p.A., Trieste, Italy.
authors.forename: Jashmini
authors.initials: J
authors.lastname: Deka
authors.affiliation: Structural Biology Laboratory, Elettra-Sincrotrone Trieste S.C.p.A., Trieste, Italy.
authors.forename: Aditya
authors.initials: A
authors.lastname: Mojumdar
authors.affiliation: Nano Innovation Laboratory, Elettra-Sincrotrone Trieste S.C.p.A., Trieste, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Parisse
authors.affiliation: Structural Biology Laboratory, Elettra-Sincrotrone Trieste S.C.p.A., Trieste, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Onesti
authors.affiliation: Nano Innovation Laboratory, Elettra-Sincrotrone Trieste S.C.p.A., Trieste, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Casalis
mesh_terms: D001483:Base Sequence; D020134:Catalytic Domain; D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D057075:Enzyme Assays / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer; D006046:Gold / Q000737:chemistry*; D057166:High-Throughput Screening Assays; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D053484:RecQ Helicases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D054852:Small Molecule Libraries; D006046:Gold; D004247:DNA; D053484:RecQ Helicases