Monitoring ligand modulation of protein-protein interactions by mass spectrometry

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Author(s) Bovet, Cedroc, Ruff, Marc, Eiler, Sylvia, Granger, Florence, Wenzel, Ryan, Nazabal, Alexis, Moras, Dino, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Monitoring ligand modulation of protein-protein interactions by mass spectrometry
Subtitle Estrogen receptor alpha-SRC1
Author(s) Bovet, Cedroc
Ruff, Marc
Eiler, Sylvia
Granger, Florence
Wenzel, Ryan
Nazabal, Alexis
Moras, Dino
Zenobi, Renato
Journal or Series Title Analytical chemistry
Volume Number 80
Issue Number 20
Start Page 7833
End Page 7839
ISSN 0003-2700
1520-6882
Publisher American Chemical Society
Publication Place Washington, DC
Publication Date 2008-00-00
Abstract Many drugs and chemicals exert their biological effect by modulating protein-protein interactions. In vitro approaches to characterize these mechanisms are often based on indirect measurements (e.g., fluorescence). Here, we used mass spectrometry (MS) to directly monitor the effect of small-molecule ligands on the binding of a coactivator peptide (SRC1) by the human estrogen receptor a ligand binding domain (hER alpha LBD). Nanoelectrospray mass spectrometry (nanoESI-MS) and high-mass matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) combined with chemical cross-linking were employed to follow these processes. The chemical cross-linking protocol used prior to high-mass MALDI analysis allows detection of intact noncovalent complexes. The binding of intact hER alpha LBD homodimer with two coactivator peptides was detected with nanoESI-MS and high-mass MALDI-MS only in the presence of an agonist ligand. Furthermore, high-mass MALDI-MS revealed an increase of the homodimer abundance after incubating the receptor with a ligand, independent of the ligand character (i.e., agonist, antagonist). The binding characteristics of the compounds tested by MS correlate very well with their biological activity reported by cell-based assays. High-mass MALDI appears to be an efficient and simple tool for directly monitoring ligand regulation mechanisms involved in protein-protein interactions. Furthermore, the combination of both MS methods allows identifying and characterizing endocrine-disrupting compounds or new drug compounds in an efficient way.
DOI 10.1021/ac8007169
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000005465
Source Database ID PP-41464
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@article{Bvt2008,
  author = "Bovet, Cedroc and Ruff, Marc and Eiler, Sylvia and Granger, Florence and Wenzel, Ryan and Nazabal, Alexis and Moras, Dino and Zenobi, Renato",
  title = "{M}onitoring ligand modulation of protein-protein interactions by mass spectrometry: {E}strogen receptor alpha-{S}{R}{C}1",
  journal = "Analytical chemistry",
  year = 2008,
  volume = "80",
  number = "20",
  pages = "7833--7839",
}


E-Citations record created: Thu, 01 Apr 2010, 23:01:50 CET