A protocol for identifying the binding sites of small molecules on the cystic fibrosis transmembrane conductance regulator (CFTR) protein

Onofrio Laselva, Evgeniy V. Petrotchenko, C. Michael Hamilton, Zafar Qureshi, Christoph H. Borchers, Robert N. Young, Christine E. Bear

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

We describe a protocol to identify the binding site(s) for a drug called ivacaftor that potentiates the CFTR chloride channel. We use photoaffinity probes—based on the structure of ivacaftor—to covalently modify the CFTR protein at the region that constitutes the drug binding site(s). We define the methods for photo-labeling CFTR, its membrane extraction, and enzymatic digestion using trypsin. We then describe the experimental methods to identify the modified peptides by using mass spectrometry. For complete details on the use and execution of this protocol, please refer to Laselva et al. (2021).

Original languageEnglish
Article number101258
JournalSTAR Protocols
Volume3
Issue number2
DOIs
Publication statusPublished - 17 Jun 2022

Keywords

  • Cell Biology
  • Cell Membrane
  • Cell-based Assays
  • Mass Spectrometry
  • Molecular/Chemical Probes
  • Protein Biochemistry

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