2. Academic Validation
  3. Proteome-Wide Identification of On- and Off-Targets of Bcl-2 Inhibitors in Native Biological Systems by Using Affinity-Based Probes (AfBPs)

Proteome-Wide Identification of On- and Off-Targets of Bcl-2 Inhibitors in Native Biological Systems by Using Affinity-Based Probes (AfBPs)

  • Chembiochem. 2018 Nov 2;19(21):2312-2320. doi: 10.1002/cbic.201800380.
Ziqian Wang 1,  Zongwei Guo 2,  Ting Song 3,  Xiaodong Zhang 3,  Nianzhe He 3,  Peng Liu 2,  Peiran Wang 3,  Zhichao Zhang 3
Affiliations

Affiliations

  • 1 Zhang Dayu School of Chemistry, State Key Laboratory of Fine Chemicals, Dalian University of Technology, No. 2 Linggong Road, Dalian, 116024, P.R. China.
  • 2 School of Life Science and Technology, Dalian University of Technology, No. 2 Linggong Road, Dalian, 116024, P.R. China.
  • 3 State Key Laboratory of Fine Chemicals, School of Chemistry, Dalian University of Technology, No. 2 Linggong Road, Dalian, 116024, P.R. China.
PMID: 30133085 DOI: 10.1002/cbic.201800380
Abstract

Selective inhibition of proteins of the Bcl-2 Family by small-molecule inhibitors is a promising new approach in drug discovery. However, information about how these molecules interact with their cellular targets (on- and off-) is highly limited. We have designed and synthesized photoreactive and "clickable" affinity-based probes (AfBPs)-Nap-2 and Nap-5-by introducing photo-crosslinkers onto Nap-1, a fluorescent derivative of small-molecule Bcl-2 Inhibitor S1-6. The resulting trifunctional probes Nap-2 and Nap-5 can enrich, visualize, and enable the identification of cellular on- and off-targets of Bcl-2 inhibitors both in vitro and in situ. Tubulin was validated as an off-target of Bcl-2 inhibitors (Nap-1 and S1-6) by large-scale cell-based proteome profiling and pull-down/western blotting (PD/WB) with Nap-2 and Nap-5. It was preliminarily illustrated to be a BH3-containing protein because some well-known Bcl-2 inhibitors can block the labeling of tubulin by Nap-2.

Keywords

Bcl-2; affinity-based protein profiling; antitumor agents; proteins; proteomics.

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