Combining Self-assembling Peptides and Proteins for Use as Templates for Nanoscale Devices

Download or read book Combining Self-assembling Peptides and Proteins for Use as Templates for Nanoscale Devices written by Kyle Webster and published by . This book was released on 2019 with total page 220 pages. Available in PDF, EPUB and Kindle. Book excerpt: Natural systems self-assemble from the nanoscale up through biomolecules, such as proteins that exhibit high functional complexity. The capabilities of proteins to control structure at the nanoscale exceed what we can achieve with top down manufacturing techniques and so can be leveraged for use as scaffolds in nanotechnology. By functionalising proteins with artificial nanostructures, we can go beyond natural biochemistry in developing new hybrid bionanotechnologies. Building on previous work, development of human peroxiredoxin three (HsPrx3) as a generalisable nanoparticle assembly system was continued. HsPrx3 was adapted through targeted mutation and protein engineering to enable functionalisation with a variety of nanoparticle cargos. HsPrx3 variants were characterised by size exclusion chromatography (SEC), transmission electron microscopy (TEM), analytical ultracentrifugation (AUC), and an X-ray crystallographic structure (Chapter 3) and tested in their capacity for unctionalisation with nanoparticle cargo. This entailed functionalisation and characterisation of HsPrx3 – gold nanoparticle conjugates followed by polyoxometelate functionalisation using a Mo132 structure (Chapter 4). Characterisation of functionalised proteins was undertaken using multiwave length SEC, spectroscopy, and energy dispersive X-ray spectroscopy (EDX). Building on previous work, demonstrating the capacity for peptide structures to template higher order HsPrx3 tube and array structures, protocols were developed for use with functionalised HsPrx3 (Chapter 5). Characterisation of HsPrx3 – peptide interactions on a SiO2 surface required combined quartz crystal microbalance with dissipation (QCM-D) and atomic force microscopy (AFM) to develop an understanding the conditions of array assembly and interactions of HsPrx3 and peptide structures with each other and a surface (Chapter 6). This work expands the array of protein scaffolds available to future researchers through design, production and testing of new HsPrx3 variants, a new X-ray crystallography structure that challenges existing understanding of the stabilising mechanism of the S78C mutation. Additionally, this research features a new HsPrx3 packing pattern and demonstrates the functionalisation of a self-assembling protein scaffold with a polyoxometalate. Finally, this is the first example of imaging and characterisation of Mo132 by TEM and EDX, verification of the protein templating capacity of two self-assembling peptide nanostructures, and the development of a novel method for validating the agreement of QCM-D and AFM data through density estimates.