title: Development of a nanoparticle-based FRET sensor for ultrasensitive detection of phytoestrogen compounds.
abstract: Phytoestrogens are plant compounds that mimic the actions of endogenous estrogens. The abundance of these chemicals in nature and their potential effects on health require the development of a convenient method to detect phytoestrogens. We have developed a nanoparticle (NP)-conjugated FRET probe based on the human estrogen receptor α (ER) ligand-binding domain (LBD) to detect phytoestrogens. The NP-conjugated FRET probe showed fluorescence signals for genistein, resveratrol and daidzein compounds with Δ ratios of 1.65, 2.60 and 1.37 respectively, which are approximately six times greater compared to individual FRET probes. A significantly higher signal for resveratrol versus genistein and daidzein indicates that the probe can differentiate between antagonistic phytoalexin substances and agonistic isoflavone compounds. NP-conjugated probes demonstrated a wide dynamic range, ranging from 10(-18) to 10(-1) M with EC(50) values of 9.6 × 10(-10), 9.0 × 10(-10) and 9.2 × 10(-10) M for genistein, daidzein and resveratrol respectively, whereas individual probes detected concentrations of 10(-13) to 10(-4) M for phytoestrogens compounds. The time profile revealed that the NP-conjugated probe is stable over 30 h and there is not a significant deviation in the FRET signal at room temperature. These data demonstrate that conjugation of a FRET probe to nanoparticles is able to serve as an effective FRET sensor for monitoring bioactive compounds with significantly increased sensitivity, dynamic range and stability.
authors.affiliation: Korea Research Institute of Biosciences and Biotechnology (KRIBB), 111 Gwahangno, Yuseong-gu, Daejeon, 305-806, Korea.
authors.forename: Arti B
authors.initials: AB
authors.lastname: Dumbrepatil
authors.forename: Seung-Goo
authors.initials: SG
authors.lastname: Lee
authors.forename: Sang J
authors.initials: SJ
authors.lastname: Chung
authors.forename: Myung Gyu
authors.initials: MG
authors.lastname: Lee
authors.forename: Byoung Chul
authors.initials: BC
authors.lastname: Park
authors.forename: Tae Jip
authors.initials: TJ
authors.lastname: Kim
authors.forename: Eui-Jeon
authors.initials: EJ
authors.lastname: Woo
mesh_terms: D001665:Binding Sites; D047628:Estrogen Receptor alpha / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D019833:Genistein / Q000032:analysis; D006801:Humans; D007529:Isoflavones / Q000032:analysis; D008024:Ligands; D053758:Nanoparticles / Q000737:chemistry*; D048789:Phytoestrogens / Q000032:analysis*; D000077185:Resveratrol; D012680:Sensitivity and Specificity; D013267:Stilbenes / Q000032:analysis
chemical_list: D047628:Estrogen Receptor alpha; D005456:Fluorescent Dyes; D007529:Isoflavones; D008024:Ligands; D048789:Phytoestrogens; D013267:Stilbenes; C004742:daidzein; D019833:Genistein; D000077185:Resveratrol