Target identification of bioactive compounds by proteomics

   Contact: Emiko Sanada  E-mail: tech_CRAST
   Collaborative Scientist: Makoto Muroi


 In response to the action of the drug, expression level or modification of proteins are changed. Using two-dimensional deference gel electrophoresis analysis system (2D-DIGE), proteome analysis of drug-treated HeLa cells is performed, and the target of the compound is estimated based on the changes in the expression data of approximately 300 spots (ChemProteoBase, Fig A). In addition, when a drug binds to its target protein, thermal stability of the protein may be changed. The change in thermal stability is also used to estimate target proteins of a compound (2DE-CETSA, Fig B).

ChemProteoBase and 2DE-CETSA

  1. Fig. A ChemProteoBase, a proteomic profiling system for drug target analysis using 2D-DIGE(1-4).

  1. Fig. B 2DE-CETSA, an analysis system for comprehensively searching the thermal stability-shifted proteins by binding with a new compound(6).


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    Methods Mol Biol, 1888: 127-139 (2019) PMID: 30519944 [ doi: 10.1007/978-1-4939-8891-4_7 ]
  5. Suvarna K, Honda K, Muroi M, Kondoh Y, Watanabe N, Osada H.: Identification of target protein for bio-active small molecule using photo-cross linked beads and MALDI-TOF mass spectrometry.
    Bio Protoc, 10(3): e3517 (2020) PMID: 33654742 [ doi: 10.21769/BioProtoc.3517 ]
  6. Nagasawa I, Muroi M, Kawatani M, Ohishi T, Ohba SI, Kawada M, Osada H.: Identification of a small compound targeting PKM2-regulated signaling using 2D gel electrophoresis-based proteome-wide CETSA.
    Cell Chem Biol, 27(2): 186-196.e4 (2020) PMID: 31813846 [ doi: 10.1016/j.chembiol.2019.11.010 ]


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