Estrogen receptor-ligand complexes measured by chip-based nanoelectrospray mass spectrometry

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Author(s) Bovet, Cédric, Wortmann, Arno, Eiler, Sylvia, Granger, Florence, Ruff, Marc, Gerrits, Bertran, Moras, Dino, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Estrogen receptor-ligand complexes measured by chip-based nanoelectrospray mass spectrometry
Subtitle An approach for the screening of endocrine disruptors
Author(s) Bovet, Cédric
Wortmann, Arno
Eiler, Sylvia
Granger, Florence
Ruff, Marc
Gerrits, Bertran
Moras, Dino
Zenobi, Renato
Journal or Series Title Protein science
Volume Number 16
Issue Number 5
Start Page 938
End Page 946
ISSN 0961-8368
Publisher Wiley
Publication Place Chichester
Publication Date 2007
Keyword(s) electrospray ionization mass spectrometry
nuclear receptor
estrogen receptor
endocrine disruptors
solution affinity
Abstract In the present report, a method based on chip-based nanoelectrospray mass spectrometry (nanoESI-MS) is described to detect noncovalent ligand binding to the human estrogen receptor α ligand-binding domain (hERα LBD). This system represents an important environmental interest, because a wide variety of molecules, known as endocrine disruptors, can bind to the estrogen receptor (ER) and induce adverse health effects in wildlife and humans. Using proper experimental conditions, the nanoESI-MS approach allowed for the detection of specific ligand interactions with hERα LBD. The relative gas-phase stability of selected hERα LBD–ligand complexes did not mirror the binding affinity in solution, a result that demonstrates the prominent role of hydrophobic contacts for stabilizing ER–ligand complexes in solution. The best approach to evaluate relative solution-binding affinity by nanoESI-MS was to perform competitive binding experiments with 17β-estradiol (E2) used as a reference ligand. Among the ligands tested, the relative binding affinity for hERα LBD measured by nanoESI-MS was 4-hydroxtamoxifen ≈ diethylstilbestrol > E2 >> genistein >> bisphenol A, consistent with the order of the binding affinities in solution. The limited reproducibility of the bound to free protein ratio measured by nanoESI-MS for this system only allowed the binding constants (Kd) to be estimated (low nanomolar range for E2). 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 technique is a promising method for screening suspected endocrine disrupting compounds and to qualitatively evaluate their binding affinity.
DOI 10.1110/ps.062664107
Additional Notes Received 15 November 2006, Revised 22 January 2007, Accepted 12 February 2007, Published Online 2 January 2009
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000627420
Source Database ID PP-34137
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  author = "Bovet, C{\'{e}}dric and Wortmann, Arno and Eiler, Sylvia and Granger, Florence and Ruff, Marc and Gerrits, Bertran and Moras, Dino and Zenobi, Renato",
  title = "{E}strogen receptor-ligand complexes measured by chip-based nanoelectrospray mass spectrometry: {A}n approach for the screening of endocrine disruptors",
  journal = "Protein science",
  year = 2007,
  volume = "16",
  number = "5",
  pages = "938--946",

E-Citations record created: Thu, 01 Apr 2010, 18:34:32 CET