Identification of endocrine-disrupting compounds using nanoelectrospray ionization mass spectrometry

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Author(s) Bovet, Cedric, Ruff, Marc, Wortmann, Arno, Eiler, Sylvia, Granger, Florence, Gerrits, Bertran, Moras, Dino, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Identification of endocrine-disrupting compounds using nanoelectrospray ionization mass spectrometry
Author(s) Bovet, Cedric
Ruff, Marc
Wortmann, Arno
Eiler, Sylvia
Granger, Florence
Gerrits, Bertran
Moras, Dino
Zenobi, Renato
Journal or Series Title Chimia
Volume Number 62
Issue Number 5
Start Page 329
End Page 334
ISSN 0009-4293
Publisher Swiss Chemical Society
Publication Place Bern
Publication Date 2008
Keyword(s) electrospray ionization mass spectrometry
endochrine-disrupting compounds
estrogen receptor
noncovalent
solution-binding affinity
Abstract A method using chip-based nanoelectrospray mass spectrometry (nanoESI-MS) is described to detect noncovalent ligand binding to the human estrogen receptor alpha ligand-binding domain (hER alpha LBD). This system represents an important environmental interest, because a wide variety of molecules, known as endocrine-disrupting compounds (EDCs), can bind to the estrogen receptor (ER) and induce adverse health effects in wildlife and humans. An efficient analytical method is therefore required to identify EDCs and characterize their solution-phase binding affinity and character (i.e. agonist or antagonist). Using proper experimental conditions, the nanoESI-MS approach allowed the detection of specific ligand interactions with hER alpha LBD. The best approach to evaluate solution-binding affinity by nanoESI-MS was to perform competitive binding experiments with 17 beta-estradiol (E2) as a reference ligand. Among the ligands tested, the relative binding affinity for hER alpha LBD measured by nanoESI-MS was 4-hydroxytamoxifen approximate to diethylstilbestrol > E2 >> genistein >> bisphenol A, consistent with the order of the binding affinities in solution. To discern agonist from antagonist, we used the specificity of a coactivator peptide for agonist-bound receptor. A specific coactivator-hER alpha LBD complex was detected only in the presence of an agonist ligand. Therefore, the specificity of nanoESI-MS combined with its speed (1 min/ligand), low sample consumption (90 pmol protein/ligand), and its sensitivity for ligand (30 ng/ml) demonstrates that this method is promising for the identification and characterization of suspected ER ligands in a high-throughput manner.
DOI 10.2533/chimia.2008.329
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 02207
03430
Organisational Unit(s)
NEBIS System Number 000000830
Source Database ID PP-41020
PP-48042
PP-41154
WOS-000256525700004
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@article{Bvt2008,
  author = "Bovet, Cedric and Ruff, Marc and Wortmann, Arno and Eiler, Sylvia and Granger, Florence and Gerrits, Bertran and Moras, Dino and Zenobi, Renato",
  title = "{I}dentification of endocrine-disrupting compounds using nanoelectrospray ionization mass spectrometry",
  journal = "Chimia",
  year = 2008,
  volume = "62",
  number = "5",
  pages = "329--334",
}


E-Citations record created: Thu, 01 Apr 2010, 22:45:19 CET