Program

Poster Presentations

Day 3, May 17（Thu.）　　Poster

3P-08　　PDF

Analysis of target proteins of bioactive compounds by using a turn-ON fluorescent affinity labeling

(¹RIKEN, ²AMED-CREST, ³JST・ERATO)
^oMiwako Asanuma^1,2,3, Takao Yamaguchi^1,3, Kosuke Dodo^1,2,3, Kenji Ohgane^1,2, Mikiko Sodeoka^1,2,3

In 2014, we reported a turn-ON fluorescent affinity labeling method, named as the O-NBD (NBD: nitrobenzoxadiazole) method (Chem. Sci., 5, 1021-1029 (2014)), for investigating target proteins of bioactive compounds. This method utilizes the property of the non-fluorescent O-NBD unit, which can be converted into a small fluorescent N-NBD unit by a covalent reaction with a proximal lysine residue. When combined with mass spectrometry, the O-NBD method makes it possible to identify the target proteins and the binding sites of bioactive compounds. As a proof-of-concept, we used biotin-avidin as a model for bioactive compound-target protein and demonstrated selective turn-ON fluorescent labeling of avidin and identified the NBD-labeled lysine residue near the biotin-binding site. Furthermore, we have applied the O-NBD method to label mouse mitochondria with an O-NBD probe for a translocator protein (TSPO). As a result, it was revealed that voltage-dependent anion channel, which is one of the partner proteins of TSPO, was labeled by O-NBD TSPO ligand.