Identification of c-di-AMP-Binding Proteins Using Magnetic Beads

Methods Mol Biol. 2017;1657:347-359. doi: 10.1007/978-1-4939-7240-1_27.


To identify cytosolic proteins that bind to cyclic di-AMP, a biotinylated analog of the nucleotide is used for protein pull-down experiments. In this approach, biotinylated c-di-AMP is coupled to Streptactin-covered beads. After protein separation using standard SDS-PAGE, the protein(s) of interest are identified by mass spectrometric analyses.

Keywords: Bacteria; Biotinylated c-di-AMP; Magnetic beads; Protein pull-down.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism
  • Biotinylation
  • DNA-Binding Proteins / metabolism*
  • Dinucleoside Phosphates / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Mass Spectrometry
  • Microspheres*
  • Protein Binding
  • Proteomics / methods


  • Bacterial Proteins
  • DNA-Binding Proteins
  • Dinucleoside Phosphates
  • cyclic diadenosine phosphate