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Title: Inhibiting protein-protein interactions in telomeres as an approach to cancer chemotherapy
Author: Salih, Twana
ISNI:       0000 0004 5920 3785
Awarding Body: University of Nottingham
Current Institution: University of Nottingham
Date of Award: 2016
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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.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QH573 Cytology ; QP501 Animal biochemistry