Studies on the Programmable Protein Assembly via Genetically Encoded Native Chemical Ligation

Abstract

Programmed protein assembly has vast potential in applications as diverse as bioreactors, smart materials and drug delivery. However, to realise this potential, exact control of the assembly process is required. Thus, this thesis describes generic genetically controlled methods to engineer the self-assembly of geometrically designed protein fusions to form user-defined structures. In particular, it shows how designed fusion proteins can be reacted to form fibres and encapsulations via split-intein mediated native chemical ligation in both one-pot and stepwise syntheses. When compatible fusions are mixed, they react quickly via a split-intein mediated native chemical ligation (NCL) to produce peptide bonded products in high yields (75 % yield). The correctly formed products can be purified to high homogeny and were shown to form the intended user-defined structures.