内源性和外源性雌激素广泛存在于食品和食品包装中,和高水平的天然雌激素和滥用或非法使用合成雌激素会导致人类内分泌紊乱,甚至癌症。因此,因此,准确评估具有雌激素样作用的食品功能性成分或毒素的存在很重要。在这项研究中,通过自组装制备了基于G蛋白偶联雌激素受体(GPERs)的电化学传感器,由双层金纳米粒子修饰,并用于测量五个GPER配体的传感动力学。17β-雌二醇传感器的互连变构常数(Ka),白藜芦醇,G-1、G-15、双酚A分别为8.90×10-17、8.35×10-16、8.00×10-15、5.01×10-15、6.65×10-16mol/L,分别。传感器对5种配体的灵敏度依次为17β-雌二醇>双酚A>白藜芦醇>G-15>G-1。受体传感器对天然雌激素的传感器灵敏度也高于外源雌激素。分子模拟对接结果表明,残基Arg,Glu,他的,GPER的Asn主要与-OH形成氢键,C-O-C,或者-NH-。在这项研究中,用电化学信号放大系统模拟细胞内受体信号级联使我们能够直接测量GPER-配体相互作用,并探索GPER在生物传感器上自组装后的动力学。本研究也为食品功能性成分和毒素的准确功能评价提供了新的平台。
Endogenous and exogenous estrogens are widely present in food and food packaging, and high levels of natural estrogens and the misuse or illegal use of synthetic estrogens can lead to endocrine disorders and even cancer in humans. Therefore, it is consequently important to accurately evaluate the presence of food-functional ingredients or toxins with estrogen-like effects. In this
study, an electrochemical sensor based on G protein-coupled estrogen receptors (GPERs) was fabricated by self-assembly, modified by double-layered gold nanoparticles, and used to measure the sensing kinetics for five GPER ligands. The interconnected allosteric constants (Ka) of the sensor for 17β-estradiol, resveratrol, G-1, G-15, and bisphenol A were 8.90 × 10-17, 8.35 × 10-16, 8.00 × 10-15, 5.01 × 10-15, and 6.65 × 10-16 mol/L, respectively. The sensitivity of the sensor for the five ligands followed the order of 17β-estradiol > bisphenol A > resveratrol > G-15 > G-1. The receptor sensor also demonstrated higher sensor sensitivity for natural estrogens than exogenous estrogens. The results of molecular simulation docking showed that the residues Arg, Glu, His, and Asn of GPER mainly formed hydrogen bonds with -OH, C-O-C, or -NH-. In this
study, simulating the intracellular receptor signaling cascade with an electrochemical signal amplification system enabled us to directly measure GPER-ligand interactions and explore the kinetics after the self-assembly of GPERs on a biosensor. This
study also provides a novel platform for the accurate functional evaluation of food-functional components and toxins.