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.

New Concepts, Experimental Approaches, and Dereplication Strategies for the Discovery of Novel Phytoestrogens from Natural Sources

“…Korean team developed a FRET biosensor probe of ERα conjugated to a nanoparticule for the detection of phytoestrogens [160]. FRET is a technique for measuring interactions between two fluorescent molecules currently used to monitor ligand-protein interactions.…”

Molecular mechanism(s) of endocrine‐disrupting chemicals and their potent oestrogenicity in diverse cells and tissues that express oestrogen receptors

“…Phyto‐oestrogens are non‐steroidal polyphenolic compounds found in some plants including soybeans, and can act as steroid hormones in animals and human beings . Diets rich in phyto‐oestrogens may exert protective effects against oestrogen‐related diseases such as ovarian and breast cancer in women although it is not clear if these anticancer effects are directly owing to the phyto‐oestrogen‐associated oestrogenicity .…”

Nanotechnology in Plant Science: To Make a Long Story Short

“…In addition, single-walled carbon nanotubes (SWNTs) have been exploited for near-infrared fluorescence monitoring of nitric oxide in A. thaliana (Giraldo et al, 2014). FRET probes conjugated to polystyrene NPs have been also designed to quantify and recognize the phytoalexins (Dumbrepatil et al, 2010).…”