title: Gold Nanoparticle-Based Förster Resonance Energy Transfer (FRET) Analysis of Estrogen Receptor: DNA Interaction.
abstract: Estrogen receptors play critical roles in regulating genes responsible for development and maintenance of reproductive tissues and other physiological function. The interaction of ERs with DNA sequences, known as estrogen response elements (EREs) (a palindromic repeat separated by three-base spacer, 5'GGTCAnnnTGACC-3'), is required for estrogen regulation of target gene expression. Here, we describe a simple "mix-and-measure"-based method for detecting ER:ERE interactions using ERE-immobilized metal nanoparticles and water-soluble conjugated polyelectrolytes (CPEs) as cooperative sensing elements. This method can differentiate the distinct DNA-binding affinity between ERα and ERβ, and determine ER:ERE-binding stoichiometry. This method can also accurately detect all 15 singly mutated EREs (i.e., three possible base substitutions at each of one to five positions from left to right of the 5' end half site, GGTCA) for their binding energy to ER. This method is compatible with 96-well plate format for high-throughput analysis.
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, 138632, Singapore. xd-su@imre.a-star.edu.sg.
authors.forename: Xiaodi
authors.initials: X
authors.lastname: Su
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, 138632, Singapore.
authors.forename: Khin Moh Moh
authors.initials: KMM
authors.lastname: Aung
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, 138632, Singapore.
authors.forename: Steven
authors.initials: S
authors.lastname: Lukman
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, 138632, Singapore.|Department of Chemical & Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore, 117576, Singapore.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
mesh_terms: D001665:Binding Sites; D002712:Chlorides / Q000737:chemistry*; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D047629:Estrogen Receptor beta / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005786:Gene Expression Regulation; D017612:Gold Compounds / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles; D009154:Mutation; D036103:Nanotechnology / Q000379:methods*; D010084:Oxidation-Reduction; D011485:Protein Binding; D020218:Response Elements; D057188:Workflow
chemical_list: D002712:Chlorides; C506487:ESR1 protein, human; C000600034:ESR2 protein, human; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D017612:Gold Compounds; C024568:gold tetrachloride, acid; D004247:DNA
keywords: Estrogen receptor; Förster resonance energy transfer; Gold nanoparticle s; Protein :DNA interaction s; Transcription factor; Water-soluble conjugated polyelectrolytes