MAPPING PROTEIN-PROTEIN INTERACTIONS BY PEPTIDE ARRAY FOR DISRUPTOR PEPTIDE DEVELOPMENT AND TRANSLATIONAL IMPACT!

Abstract

Aberrant cellular signalling is central to the pathogenesis of a number of neurodegenerative disorders. Our group focuses in particular on the role of Wnt signalling in β-amyloid toxicity, and have demonstrated not only that aberrant Wnt activation, APP proteolysis and synapse loss are intimately linked, but have identified the key protein-protein interactions responsible. A fundamental aspect of our pathway-driven approach is the mapping and disruption of protein-protein interactions using peptide array technology. Peptide arrays are created from peptide libraries consisting of overlapping peptides spanning the entire sequence of the protein of interest and spotted onto cellulose coated slides. These peptide arrays can be overlaid with putative binding partners to identify potential binding regions or novel post-translational modification sites. Key amino acid residues can be rapidly identified by alanine/ single substitution scanning. Crucially, we can delineate the minimal “core” binding regions, which can be synthesised and used as cell permeable “disruptor peptides” capable of selectively blocking specific protein-protein interactions. Furthermore, disruptor peptides can be used as the basis of high throughput biochemical and cell based assays for drug discovery. We wish to showcase the power of this technology, using specific examples from our own work on Alzheimer's disease, and demonstrate its capabilities, which are readily amenable across a broad range of applications, to provide useful research tools in dissecting specific signalling events. Mapping and disrupting protein-protein interactions by peptide array has significant potential to provide translational outcomes for neurodegenerative disease.