Characterization of the binding between phthalate esters and mouse PPARα for the development of a fluorescence polarization-based competitive binding assay

Zhang, Jie; Xing, XiaoJia; Sun, Yonghai; Li, Zhuolin; Xue, Peiyu; Wang, Tuoyi; Li, Tiezhu

doi:10.1039/c5ay03053f

Cited by 13 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Based on the structural similarity of ginsenosides to steroid hormones, some pharmacological effects of ginsenosides may be mediated by binding to nuclear receptors, such as androgen receptor, estrogen receptors (ERs), glucocorticoid receptor, peroxisome proliferator-activated receptor γ, and progesterone receptor [9] , [10] , [11] , [12] . Nuclear receptor superfamily is a group of transcription factors that can be activated by the functional ligands, including 17β-estradiol, dexamethasone, fatty acids, etc [13] , [14] , [15] . As the main targets of phytoestrogens, ERs have been reported for their ability of binding several ginsenosides, including Re and Rh 1 [9] , [16] .…”

Section: Introductionmentioning

confidence: 99%

Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

Zhang

Zhong

Hou

et al. 2020

Journal of Ginseng Research

Self Cite

View full text Add to dashboard Cite

Background As the main metabolites of ginsenosides, 20( S , R )-protopanaxadiol [PPD( S , R )] and 20( S , R )-protopanaxatriol [PPT( S , R )] are the structural basis response to a series of pharmacological effects of their parent components. Although the estrogenicity of several ginsenosides has been confirmed, however, the underlying mechanisms of their estrogenic effects are still largely unclear. In this work, PPD( S , R ) and PPT( S , R ) were assessed for their ability to bind and activate human estrogen receptor α (hERα) by a combination of in vitro and in silico analysis. Methods The recombinant hERα ligand-binding domain (hERα-LBD) was expressed in E. coli strain. The direct binding interactions of ginsenosides with hERα-LBD and their ERα agonistic potency were investigated by fluorescence polarization and reporter gene assays, respectively. Then, molecular dynamics simulations were carried out to simulate the binding modes between ginsenosides and hERα-LBD to reveal the structural basis for their agonist activities toward receptor. Results Fluorescence polarization assay revealed that PPD( S , R ) and PPT( S , R ) could bind to hERα-LBD with moderate affinities. In the dual luciferase reporter assay using transiently transfected MCF-7 cells, PPD( S , R ) and PPT( S , R ) acted as agonists of hERα. Molecular docking results showed that these ginsenosides adopted an agonist conformation in the flexible hydrophobic ligand-binding pocket. The stereostructure of C-20 hydroxyl group and the presence of C-6 hydroxyl group exerted significant influence on the hydrogen bond network and steric hindrance, respectively. Conclusion This work may provide insight into the chemical and pharmacological screening of novel therapeutic agents from ginsenosides.

show abstract

Section: Introductionmentioning

confidence: 99%

Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

Zhang

Zhong

Hou

et al. 2020

Journal of Ginseng Research

Self Cite

View full text Add to dashboard Cite

show abstract

“…Fluorescence polarization (FP) assay is founded on the exceptional specificity of antibodies for their target antigens and has been extensively employed in clinical diagnostics . Nowadays, the receptor‐based FP assay has already been successfully applied to the detection of ochratoxin A , 17β‐estradiol , ractopamine , and phthalate esters . As a simple and rapid method, broad‐specific receptor‐based FP assay has several advantages, such as short assay time and absence of washing step in comparison with the enzyme‐linked immunosorbent assay.…”

Section: Introductionmentioning

confidence: 99%

Estrogen receptor‐based multi‐residue screening of bisphenol compounds in urine

Guan

Sun

et al. 2018

Biotech and App Biochem

Self Cite

View full text Add to dashboard Cite

Human exposure to bisphenol compounds (BPs) has been implicated in the development of several chronic diseases. Instead of exploiting the traditional methods for determination of BPs, this work confirms that the human estrogen receptor α ligand binding domain (hERα-LBD) is a powerful recognition element that can be used to monitor multi-residue of BPs in urine samples by fluorescence polarization (FP) assay. Test parameters were optimized for the best performance. Under the optimal conditions, the IC 50 values of BPs are in the range of 0.04-1.61 μg mL −1 . Recovery experiments were then performed to assess the accuracy and precision of the established method. The results detected by FP assay show good agreements with that of liquid chromatography-tandem mass spectrometry method with a fit of R 2 = 0.9372 and 0.9640 for BPE and BPAP, respectively. A computational methodology, ligand-based pharmacophore model, was also employed to further explore the broad-specific of tested compounds. It was found that the two hydrogen bond acceptor features and one hydrophobic aliphatic feature were essential for the corresponding cross-reactivity results from the FP assay. All these results suggest that the established method can be successfully applied to monitor the occurrence

show abstract

“…[19] Moreover, PPARα has been reported to bind to an even wider range of ligands than either PPARβ or PPARγ. [20] Lines of evidence showed that PPARα could be activated by BPs. [21,22] The objective of this work was to build a competitive FP assay based on the mouse peroxisome proliferator-activated receptor α ligand binding domain (mPPARα-LBD*) for simultaneous monitoring BPA, BPF, BADGE, and BFDGE.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence polarization assay for the simultaneous determination of bisphenol A, bisphenol F and their diglycidyl ethers in canned tuna

Guan

Zhang

et al. 2017

International Journal of Food Properties

Self Cite

View full text Add to dashboard Cite

This work developed a fluorescence polarization (FP) assay for simultaneous monitoring BPA, BPF, BADGE, and BFDGE in canned tuna. The assay was based on the competitive binding between bisphenol analogues (BPs) and dexamethasone fluorescein (Dex-fl) for mouse peroxisome proliferator-activated receptor α ligand binding domain (mPPARα-LBD*). The soluble form of mPPARα-LBD* was expressed in Escherichia coli strain Rosetta (DE3), as a recognition element on detection of BPs. Under optimized conditions, four bisphenol analytes were detected at concentrations corresponding to the specific migration limits (SMLs) as required by the European Union. The FP assay showed IC 50 values of 2.80, 6.51, 1.72, and 4.28 mg L −1 with a limit of detection of 0.35, 0.08, 0.10, and 0.49 mg L −1 for BPA, BPF, BADGE, and BFDGE, respectively. The analysis of spiked canned tuna samples showed the acceptable recoveries ranged from 87.9% to 77.3%, with variation coefficients ranging from 9.0% to 15.8%. With the high sensitivity and wide-range affinities to BPs, the developed assay based on mPPARα-LBD* exhibited the potential to be a screening assay for fast detection of BPs in canned tuna.

show abstract

Characterization of the binding between phthalate esters and mouse PPARα for the development of a fluorescence polarization-based competitive binding assay

Abstract: The binding between PPAR and PAEs was investigated for developing an efficient fluorescence polarization-based competitive binding assay.

Cited by 13 publications

References 33 publications

Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

Estrogen receptor‐based multi‐residue screening of bisphenol compounds in urine

Fluorescence polarization assay for the simultaneous determination of bisphenol A, bisphenol F and their diglycidyl ethers in canned tuna

Contact Info

Product

Resources

About