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This article aims to initial screening through the use of molecular docking technology may receptor hERα hERRγ widely used hVDR combination of food and the presence of potential EDCs, and then build hERα hERRγ hVDR three recombinant plasmids, prokaryotic expression hERα, hERRγ hVDR three receptor protein, set up to explore the potential EDCs recombinant receptor protein binding activity of phosphate nitrophenol food, explore may hERα hERRγ the hVDR receptor channel on the human body harmful substances. Gene, cloned from H295R cells hERRγ the hVDR gene from Caco-2 cell clones were constructed recombinant plasmid pGEX-4T-1-hERRγ, pGEX-4T-1-hVDR. The recombinant plasmid was transformed into E. coli DH5α, after the blue-white screening digestion verify sequencing prove hERRα such hVDR has been successfully cloned and transformed. Prokaryotic expression and purification of GST-hERRγ, the GST-hVDR fusion to the protein expression and purification laboratory has constructed a recombinant strain pGEX-4T-1-hERα, the pET28-TIF2-BAP-'s GST-hERα, TIF2-BAP fusion protein . HERα nitro-phenol method, hERRγ binding activity of the phosphoric acid p-nitro phenol method, hVDR binding activity of phosphate binding activity of the phosphate nitrophenol. Molecular docking technology, initial screening may have hERα hERRγ a hVDR combined potential EDCs widespread activity in six categories of food were: methylparaben class, salicylic acid esters, phthalic esters, bisphenol A and its analogues, isoflavones, heavy metals. Binding activity of the phosphorylated recombinant receptor protein nitro-phenol method, determination of the p-hydroxybenzoic acid esters, salicylic acid esters, phthalate esters, bisphenol A and its analogues, isoflavones, heavy metals the class six categories of substances with receptor hERα hERRγ hVDR the binding activity, and conducted the comparative analysis. The results show that the hydroxy benzoic acid esters and hERα hERRγ have binding activity, salicylic acid esters, and other salicylic acid esters in addition to virtually no binding activity of phenyl salicylate with hERRγ with hERα hERRγ binding activity. Phthalate substances have hERα binding activity but only weak activity dimethyl phthalate and diethyl phthalate and hERRγ, EC50 were 2.77 × 10 4 mol / L, 3.07 × 10 -3mol / L. Of bisphenol A and its analogues, 4,4 - biphenyldiol hERα has a strong binding activity, EC50 of 6.80 x 10-6mol / L, tetrabromobisphenol A with hERRα only binding activity, EC50 achieve 3.11 × 10-5mol / L. The isoflavones, heavy metal substances hERα, hERRγ there is a certain activity. the hVDR receptor active only with heavy metals. The use of molecular docking simulation technology, using recombinant receptor protein binding activity phosphate measured nitrophenol typical EDCs for further scoring authentication, results show that the docking simulation results based on the protein binding activity of recombinant receptor phosphorylation on The nitrophenol Determination consistent.
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