Inhibiting protein–protein interactions in telomeres as an approach to cancer chemotherapy

Salih, Twana (2016) Inhibiting protein–protein interactions in telomeres as an approach to cancer chemotherapy. PhD thesis, University of Nottingham.

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Stable telomeres play a key role to the survival of cancer cells; therefore, different cancer chemotherapeutic approaches have been developed in order to disrupt or destabilise telomeres or telomerase. One of the newest methods is the disruption of vital protein–protein interactions in the telomere, such as that between shelterin components TRF1 and TIN2. The principal aim of this project was to obtain a novel peptide-like molecule, an analogue of a key interacting region of TIN2 that could compete effectively for the binding sites on TRF1 and so lead to the destabilisation of telomere structure. Molecular modelling and simulations were undertaken as the starting point of the project. Structure-based drug design was applied, starting from the available crystal structure data. A library of peptide analogues of the TRF1-binding motif in TIN2 was designed using the MM-GBSA simulation method to predict binding affinities. Then, a number of the peptide analogues were selected from the library for further investigations.

The secondary goal was to investigate the accuracy of the predicted ΔGbinding values and try to optimise them; the latter aim was set out after finding a significant difference in the predicted binding free energy values after repeating the identical protocol for the same complex system. Therefore, different approaches were applied to optimise the predicted ΔGbinding values. Subsequently, selected TIN2 peptide analogues were synthesised in the laboratory using Fmoc solid-phase peptide synthesis. Then, the hTRF1 protein was expressed and purified in preparation for the development of the in vitro assay. Finally, biophysical evaluations and screening of the peptide analogues were performed using fluorescence polarisation assay.

One of the peptide analogues developed in this study was identified as an early lead compound. In addition, the findings of this research showed that the ΔGbinding values of the peptide analogues have significantly improved accuracy after optimisation. As a result of these investigations, suggestions were identified for future research.

Item Type: Thesis (University of Nottingham only) (PhD)
Supervisors: Laughton, Charles
Chan, Weng
Subjects: Q Science > QH Natural history. Biology > QH573 Cytology
Q Science > QP Physiology > QP501 Animal biochemistry
Faculties/Schools: UK Campuses > Faculty of Science > School of Pharmacy
Item ID: 32001
Depositing User: Salih, Twana
Date Deposited: 28 Jul 2016 13:27
Last Modified: 01 Oct 2016 09:55

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