The yeast prion protein Ure2 consists of an N-terminal prion-inducing domain and a C-terminal functional domain. The prion properties of Ure2 are determined by the N-terminal prion domain (PrD), which contains key sequence regions that drive amyloid fibril assembly; the C-terminal domain shows enzymatic activities which are dependent upon the dimeric arrangement of C-domains in both the soluble and fibrillar form of Ure2. Self-assembly is one of a number of features of amyloid fibrils that make them attractive targets for biotechnology. The purpose of this research is to generate novel protein-based assemblies for use in bio-/nanotechnology, using the Ure2-based amyloid fibrils as a scaffold for the display of bioactive macromolecules. We will test whether other dimeric enzymes, such as Alkaline phsophatase (AP) and Horseradish peroxidase (HRP), can be supported in functional conformations on Ure2 fibrils. ..First, we will employ standard molecular biology techniques to generate a range of Ure2-linker-AP/HRP fusion genes, which will be then overexpressed in Escherichia coli. The next step will be to determine the conditions necessary for fibril assembly, involving screening fibril material against a variety of conditions (pH, ionic strength, buffer components). The amyloid fibrils formed from the designed fusion sequences will be characterised by Transmission Electron Microscopy (TEM), X-ray fibre diffraction and by solution structure techniques such as Circular Dichroism (CD) and Fourier Transform Infrared (FTIR) spectroscopy. Finally, the amount of active enzyme displayed on the fibril surface can be determined using a well-characterized colorimetric assay. By comparing the enzymatic activities of fibrils comprising different combinations of Ure2 and linker sequences we will develop models describing how the proportion of active enzyme varies with the length of the amyloidogenic and linker peptides and the arrangement of monomers and dimers within the fibril. ..This research will enhance our understanding of the molecular processes underlying amyloid fibril structure and assembly, which may lead to the development of therapeutics to prevent or treat protein misfolding diseases such as Alzheimer’s disease. This research will also enable us to determine the parameters required for the conformationally active display of a variety of enzymes, and thereby achieve great control over the display process itself. This research also has important implications for the development of efficient and robust enzyme immobilisation technologies, which is of vast economic importance in industrial processes that employ biocatalysts.