Precise, autocatalytic chemistry on proteins and peptides

This project will investigate site-specific acylation of proteins and peptides, with a focus on developing improved therapeutics. Different functional moieties will be attached to proteins or peptides via optimized amino acid sequences (Lys-His tag) placed in flexible terminals or internal loop regions to achieve or enhance serum-stability and/or cell-toxicity. The central focus will be on the protein alpha-1 antitrypsin (A1AT), belonging to the serpin superfamily and the antibody Rituximab. Proteins comprising human-like glycosylation patterns will be achieved by production in glyco-engineered Saccharomyces cerevisiae and Chinese Hamster Ovary cells. Furthermore, peptide inhibitors of the essential fungal protein Pma1 will be modified to enhance serum-stability and cell-permeability. Lys-His tags will be optimized to achieve optimal acylation efficiency, and the activity and stability of acylated protein/peptide conjugates will be assessed through binding studies, enzyme assays, and cell-toxicity assays.