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

STAR Protocols

Christine E. Bear¹  C. Michael Hamilton¹  Christoph H. Borchers²  Zafar Qureshi²  Onofrio Laselva³  Evgeniy V. Petrotchenko⁴  Robert N. Young⁴ 

[1] Center for Computational and Data-Intensive Science and Engineering, Skolkovo Institute of Science and Technology, Moscow, Russia;Department of Chemistry, Simon Fraser University, 8888 University Dr., Burnaby, BC V5A 1S6, Canada;Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy;Segal Cancer Proteomics Center, Lady Davis Institute, Jewish General Hospital, McGill University, Montreal, QC, Canada;

关键词: Cell Biology;    Cell Membrane;    Cell-based Assays;    Molecular/Chemical Probes;    Protein Biochemistry;    Mass Spectrometry;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown