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Title: Exploring solid state NMR spectroscopy as a tool to study F-actin and its complexes
Author: Pace, Nicola
ISNI:       0000 0004 6347 9960
Awarding Body: King's College London
Current Institution: King's College London (University of London)
Date of Award: 2017
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Actin plays a central role in eukaryotic cells in various physiological processes, such as cell motility, cell division and DNA transcription. In addition, actin is the key protein in cardiac muscles, forming filaments that are crucial to its function. The possibility to have the structure of these actin filaments at atomic resolutions is fundamental to understand the molecular mechanism of these events. Although the atomic structure of the monomer of actin has been already characterized, only models based on data at low resolution are available for the structure of actin filaments. This is mainly due to the impossibility of exploiting the full potential of the two most common techniques used in structural biology: NMR in solution and x-ray crystallography. The former cannot be used because the size of actin exceeds the limit of detection, the latter because of the intrinsically disorder contained in a sample of actin filaments which does not allow the growing of a useful crystal. This work aims to investigate the possible application of solid state NMR spectroscopy to the study of actin filament and its complexes, as this technique is not affected by the above mentioned limitations. In order to simplify the approach to this technique we decided to start with the study of a small actin binding domain, the Villin Head Domain (VHD), complexed with F-actin. Despite the limited number of VHD residues (67), its assignment in the bound state is not still complete, meaning that the structure of the complex is not available yet. Nevertheless, thanks to the implementation of phosphorous and Double-Quantum ssNMR spectroscopy, alongside the possibility to acquire proton spectra at high resolution, it has been possible to create a platform of experiments that can be used to approach the study of thin filament complexes in order to solve their structure. The availability of the structure of F-actin and its complexes at atomic resolution will be the starting point for further investigations to understand how actin is implicated in health and disease.
Supervisor: Pfuhl, Mark Christoph Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available