Characterizing the protein-protein interaction between MDM2 and 14-3-3σ; proof of concept for small molecule stabilization

被引:4
作者
Ward, Jake A. [1 ,2 ]
Romartinez-Alonso, Beatriz [1 ,3 ]
Kay, Danielle F. [4 ]
Bellamy-Carter, Jeddidiah [4 ]
Thurairajah, Bethany [5 ]
Basran, Jaswir
Kwon, Hanna
Leney, Aneika C. [4 ]
Macip, Salvador [6 ,7 ]
Roversi, Pietro [1 ,8 ]
Muskett, Frederick W. [1 ]
Doveston, Richard G. [1 ,5 ]
机构
[1] Leicester Inst Struct & Chem Biol, Leicester, England
[2] Mech Canc & Aging Lab, Dept Mol & Cell Biol, Leicester, England
[3] Univ Leicester, Dept Mol & Cell Biol, Leicester, England
[4] Univ Birmingham, Sch Biosci, Birmingham, England
[5] Univ Leicester, Sch Chem, Leicester, England
[6] Univ Oberta Catalunya, Fac Hlth Sci, FoodLab, Barcelona, Spain
[7] Josep Carreras Leukaemia Res Inst, Cami Escoles S-N, Badalona, Barcelona, Spain
[8] Inst Agr Biol & Biotechnol, Unit Milan, Milan, Italy
基金
英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会;
关键词
BINDING; P53; ACTIVATION; DOMAIN;
D O I
10.1016/j.jbc.2024.105651
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mouse Double Minute 2 (MDM2) is a key negative regulator of the tumor suppressor protein p53. MDM2 overexpression occurs in many types of cancer and results in the suppression of WT p53. The 14-3-3 family of adaptor proteins are known to bind MDM2 and the 14-3-3o isoform controls MDM2 cellular localization and stability to inhibit its activity. Therefore, small molecule stabilization of the 14-3-3o/MDM2 protein-protein interaction (PPI) is a potential therapeutic strategy for the treatment of cancer. Here, we provide a detailed biophysical and structural characterization of the phosphorylationdependent interaction between 14-3-3o and peptides that mimic the 14-3-3 binding motifs within MDM2. The data show that di-phosphorylation of MDM2 at S166 and S186 is essential for high affinity 14-3-3 binding and that the binary complex formed involves one MDM2 di-phosphorylated peptide bound to a dimer of 14-3-3o. However, the two phosphorylation sites do not simultaneously interact so as to bridge the 14-3-3 dimer in a ' multivalent ' fashion. Instead, the two phosphorylated MDM2 motifs ' rock ' between the two binding grooves of the dimer, which is unusual in the context of 14-3-3 proteins. In addition, we show that the 14-3-3o-MDM2 interaction is amenable to small molecule stabilization. The natural product fusicoccin A forms a ternary complex with a 14-3-3o dimer and an MDM2 di-phosphorylated peptide resulting in the stabilization of the 14-3-3o/MDM2 PPI. This work serves as a proof- of-concept of the drugability of the 14-3-3/MDM2 PPI and paves the way toward the development of more selective and efficacious small molecule stabilizers.
引用
收藏
页数:13
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