Differential binding with ERα and ERβ of the phytoestrogen-rich plant Pueraria mirifica

Pueraria mirifica leaves, an alternative potential isoflavonoid source

“…15) The sensitivity of all tested compounds determined by YES-hERαΔsnq2 and YES-hERβΔsnq2 was higher than the corresponding wildtype Y190 yeast strains (YES-hERα and YES-hERβ), which correlated well with a previous study. 33) In wildtype Y190 yeast strains, anti-estrogenic activity could be determined by using tamoxifen against both E 2 34) and P. mirifica tuber crude extract.…”

“…For the YES-hERβ assay, Y190 was transformed with pGBT9-hERβLBD, encoding for amino acid residues 213-477 of the hER β fused to GAL4DBD, and pGAD424-hSRC1, encoding for amino acid residues 231-1094 of steroid receptor co-activator 1 (SRC1) fused to GAL4AD. 15) In addition, the YES-hERαΔsnq2 and YES-hERβΔsnq2 systems were constructed from the YES-hERα and YES-hERβ systems as above except using the Y190-Δsnq2 as the host strain in place of the wildtype Y190 so as to increase the sensitivity of YES system. Plant material.…”

“…The methodology was described in previous study. 15) A stock solution of each crude leafextract was freshly prepared as serial dilutions (50-2,000 μg/mL) in DMSO. Standard compounds: E 2 , genistein, and genistin were also prepared in DMSO in dose ranges of 10 −16 -10 −4 M, 10…”

“…The readout is then achieved by performing a β-galactosidase assay through the conversion of oNPG to oNP, which is monitored by absorbance of the oNP at 420 nm and compared to that for E 2 as a positive control and reference standard. 15) For the standardization of the YES-hER assay with the wildtype and Δsnq2 yeast strains for the evaluation of estrogenic activity, the EC 50 values of E 2 in the YES-hERα and YES-hERβ assays (Table 2) were found to agree fairly well (1.2-fold lower activity) with that previously reported for YES-hERα (2.25 × 10 −10 M), but was some 51.3-fold less active than that reported before for YES-hERβ (2.3 × 10 −10 M). In this study here, the YES-hERα exhibited a 44.2-fold higher estrogenic activity than the YES-hERβ system, whilst the YES-hERαΔsnq2 assay exhibited a 283.6-fold higher estrogenic activity than that in the YES-hERβΔsnq2 system.…”

“…11) Pueraria mirifica Airy Shaw et Suvatabhandu is a Thai indigenous legume herb with a long-term folk medicinal consumption among Thai women for menopausal treatment. The tuberous materials of the plant revealed strong estrogenic effects in the MCF-7 proliferation/antiproliferation, 12) uterotrophic 13,14) and Yeast Estrogen Screen (YES) 15) test assays. Among the phytoestrogen-rich plant materials, the tubers of kudzu (Pueraria lobata) are widely used in traditional Chinese, Japanese, and Korean medicines, 16) whereas in traditional Thai medicines the related P. mirifica is used and has been extensively studied.…”