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Updated: Dec 26, 2025
A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
Published on: April 29, 2022
Identification of Phosphate-Containing Compounds as New Inhibitors of 14-3-3/c-Abl Protein-Protein Interaction
Leire Iralde-Lorente1, Giusy Tassone1, Letizia Clementi2
1Department of Biotechnology, Chemistry and Pharmacy-Department of Excellence 2018-2022, Università degli Studi di Siena, via Aldo Moro 2, 53100 Siena, Italy.
Abstract:
The 14-3-3/c-Abl protein-protein interaction (PPI) is related to carcinogenesis and in particular to pathogenesis of chronic myeloid leukemia (CML). Previous studies have demonstrated that molecules able to disrupt this interaction improve the nuclear translocation of c-Abl, inducing apoptosis in leukemia cells. Through an X-ray crystallography screening program, we have identified two phosphate-containing compounds, inosine monophosphate (IMP) and pyridoxal phosphate (PLP), as binders of human 14-3-3σ, by targeting the protein amphipathic groove. Interestingly, they also act as weak inhibitors of the 14-3-3/c-Abl PPI, demonstrated by NMR, SPR, and FP data. A 37-compound library of PLP and IMP analogues was investigated using a FP assay, leading to the identification of three further molecules acting as weak inhibitors of the 14-3-3/c-Abl complex formation. The antiproliferative activity of IMP, PLP, and the three derivatives was tested against K-562 cells, showing that the parent compounds had the most pronounced effect on tumor cells. PLP and IMP were also effective in promoting the c-Abl nuclear translocation in c-Abl overexpressing cells. Further, these compounds demonstrated low cytotoxicity on human Hs27 fibroblasts. In conclusion, our data suggest that 14-3-3σ targeting compounds represent promising hits for further development of drugs against c-Abl-dependent cancers.
Insights
Two phosphate compounds, inosine monophosphate (IMP) and pyridoxal phosphate (PLP), were identified as inhibitors of the 14-3-3/c-Abl protein-protein interaction. These compounds show promise for developing new drugs against chronic myeloid leukemia (CML).
Area of Science:
- Biochemistry and Molecular Biology
- Cancer Research
- Drug Discovery
Background:
- The 14-3-3/c-Abl protein-protein interaction (PPI) is implicated in carcinogenesis, particularly in chronic myeloid leukemia (CML).
- Disrupting this PPI can enhance c-Abl nuclear translocation, leading to apoptosis in leukemia cells.
Purpose of the Study:
- To identify novel small molecules that can disrupt the 14-3-3/c-Abl PPI.
- To evaluate the therapeutic potential of identified compounds against c-Abl-dependent cancers.
Main Methods:
- X-ray crystallography screening identified inosine monophosphate (IMP) and pyridoxal phosphate (PLP) as binders of human 14-3-3σ.
- Nuclear Magnetic Resonance (NMR), Surface Plasmon Resonance (SPR), and Fluorescence Polarization (FP) assays confirmed weak inhibition of the 14-3-3/c-Abl PPI.
- Antiproliferative activity was assessed against K-562 leukemia cells, and c-Abl nuclear translocation was measured.
Main Results:
- IMP and PLP were identified as weak inhibitors of the 14-3-3/c-Abl PPI, targeting the amphipathic groove of 14-3-3σ.
- A library of analogues yielded three additional weak inhibitors.
- IMP and PLP demonstrated significant antiproliferative effects on K-562 cells and promoted c-Abl nuclear translocation with low cytotoxicity to fibroblasts.
Conclusions:
- IMP and PLP are promising lead compounds for developing novel therapeutics targeting c-Abl-dependent cancers.
- Targeting the 14-3-3σ protein offers a viable strategy for CML treatment.
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