کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2602799 | 1133798 | 2009 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Towards high-throughput identification of endocrine disrupting compounds with mass spectrometry Towards high-throughput identification of endocrine disrupting compounds with mass spectrometry](/preview/png/2602799.png)
High-mass matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) combined with chemical cross-linking has the ability to monitor the ligand-dependent dimerization of the human estrogen receptor α ligand binding domain (hERα LBD) in solution. Because only ER ligands enhance the homodimer abundance, we evaluated the ability of this label-free approach for identifying endocrine disrupting compounds (EDCs) in a high-throughput manner. This was achieved by combining an automated liquid handler with an automated MS acquisition procedure, which allowed a five-fold gain in operator time compared to a fully manual approach. To detect ligand binding with enough confidence, the receptor has to be incubated with at least a 10 μM concentration of the test compound. Based on the increase of the measured homodimer intensity, eight compounds with a relative binding affinity (RBA, relative to the natural hormone estradiol) >7% were identified as ER ligands among the 28 chemicals tested. Two other compounds, quercetin and 4-tert-amylphenol, were also identified as ER ligands, although their RBAs have been reported to be only 0.01% and 0.000055%, respectively. This suggests that these two ligands have a higher affinity for hERα LBD than reported in the literature. The high-mass MALDI approach thus allows identifying high affinity EDCs in an efficient way.
Journal: Toxicology in Vitro - Volume 23, Issue 4, June 2009, Pages 704–709