CEEPC/IPM/CMSC - Abstrakt prezentace

(CEEPC/IPM/CMSC 2022 - ThP-33)
HDX-MS study of the HDM2 protein interaction with the Nutlin-3 compared to p53 protein

Josef Kucera 1 *, Lucia Haronikova 1, Lukas Uhrik 1, Ondrej Bonczek 1, Tomas Henek 1, Vaclav Hrabal 1, Lixiao Wang 2, Borivoj Vojtesek 1, Robin Fahraeus 1,3, Lenka Hernychova 1

  1. RECAMO, Masaryk Memorial Cancer Institute, Zluty kopec 7, 602 00 Brno, Czech Republic
  2. Department of Medical Biosciences, Umea University, 901 87 Umea, Vasterbotten, Sweden
  3. Inserm UMRS1131, Institut de Gén. Mol., Université Paris 7, Hôpital St. Louis, 750 10 Paris, France

Abstrakt

Cancer is defined as uncontrolled cell division and tumour cell growth with the potential to spread or invade other body organs. The p53 protein, known as the guardian of the genome, plays important role in tumour suppression. Its interaction with human double minute (HDM2) inhibits p53 transcriptional activity and stimulates its ubiquitination followed by degradation. Overexpression of HDM2 in tumours inhibits p53 and therefore promotes uncontrolled cell proliferation. Inhibition of this interaction is important for cancer therapy. One of the promising inhibitors is Nutlin-3 (cis-imidazoline), which interacts with the HDM2 at the p53 binding site, thereby able to disrupt the p53 HDM2 complex. This leads to the release and activation of p53 and consequent cell cycle arrest or apoptosis.
In this work, hydrogen-deuterium exchange mass spectrometry (HDX MS) was employed for the study of HDM2 conformational dynamics after binding either with Nutlin-3 or p53 protein. The higher suppression of deuteration at the p53 binding domain in HDM2 protein was observed after the Nutlin-3 binding compared to the p53-HDM2 complex. Unlike p53-HDM2 interaction, the Nutlin-3 induced the allosteric changes in the C-terminal disordered region of the HDM2.
The results show that Nutlin-3 mimics the HDM2 binding site of p53. The different HDX suppression was observed in the case of Nutlin-3 interaction in comparison to p53 binding to HDM2. This pilot study will continue further with small molecule inhibitors which have the potential for cancer treatment.

* Korespondující autor: josef.kucera@mou.cz

Literatura

  1. Haronikova, L. et al.: Cell. Mol. Biol. Lett. 26 (1), (2021)
  2. Hernychova, L. et al.: Proteomics 13 (16), 2512-2525 (2013)
  3. Li, R. et al.: Cell. Mol. Biol. Lett. 25 (1), (2020)

Poděkování:

Supported by the European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868), MH CZ - DRO (MMCI, 00209805), National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU and Czech Science Foundation project (No. 19-18177Y).


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