In Silico Design of Protein–Protein Interaction Libraries for Fasciculin-2 Variant Selection

The interaction between the toxin fasciculin-2 (Fas-2) and the enzyme acetylcholinesterase (AChE) represents a fundamental model for studying protein-protein interactions, particularly in mechanisms associated with neurodegenerative diseases such as Alzheimer and other conditions associated with synaptic dysfunction. The aim of this project is to use structural bioinformatics tools to guide the design of a Fas-2 mutant library, to be screened by Yeast Surface Display (YSD), with the purpose of identifying variants with lower toxicity. The methodology combines structural bioinformatics and molecular modeling using the Rosetta software suite to predict the effects of mutations on stability and binding interfaces. This computational approach also allows for the rational selection of priority targets for more in-depth biophysical studies, such as thermal stability measurements, conformational dynamics, and structural assays via protein crystallography. The expected results will not only advance the understanding of protein–protein interactions in this system but also contribute to the development of biotechnological and medical applications by providing rationally engineered, less toxic protein variants.