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Peroxisome Proliferator-Activated Receptors (PPARs)

Volume 952 of the series Methods in Molecular Biology pp 287-299

Date:

Analyzing PPARα/Ligand Interactions by Chemical Cross-Linking and High-Resolution Mass Spectrometry

  • Mathias Q. MüllerAffiliated withDepartment of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin-Luther-Universität Halle-Wittenberg
  • , Christian H. IhlingAffiliated withDepartment of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin-Luther-Universität Halle-Wittenberg
  • , Andrea SinzAffiliated withDepartment of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin-Luther-Universität Halle-Wittenberg Email author 

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Abstract

The combination of chemical cross-linking and high-resolution mass spectrometry is an emerging technique for monitoring conformational changes in proteins induced by drug binding. In this chapter, we describe this approach for gaining insights into the conformational changes of the peroxisome proliferator-activated receptor alpha after binding of low-molecular weight ligands. Our strategy provides a basis to efficiently characterize target protein-drug interactions.

Key words

PPARα/ligand interaction Chemical cross-linking Isotope labeling SDS-PAGE In-gel digestion Nano-HPLC High-resolution mass spectrometry