Executive Summary : | This proposal aims to expand the toolbox of biologically useful optical probes by studying a palette of noncanonical amino acids (NcAAs) with different excitation and emission regions. A series of FRET (Förster resonance energy transfer) pairs will be designed for the further development of stimuli responsive noncanonical peptide-based optical probes (NcPOPs) targeting important biomarkers. The scope of the project will be extended by synthesizing new and literature-reported NcAAs, which will be used to generate NcPOPs for biomarker detection. A simple thiol responsive screening platform will be proposed for rapid screening of NcAA-based FRET pairs, allowing for fluorescence "donor" and "acceptor" type NcAAs to be monitored in the absence and presence of a thiol trigger. The advantage of this assay platform lies in its simple design and mode of operation to monitor FRET efficiency between the fluorescence donors and acceptors. In the final stage of the proposal, various NcPOPs targeting biological markers will be designed and synthesized. These NcPOPs will be made varying peptide chain sequence and fluorogenic or quenchergenic NCAAs to optimize efficiency. The proposal targets two important biomarkers, histone deacetylase (HDAC) enzyme isoforms and S-nitrosylated proteins. The major advantage of these NcPOPs lies in their flexibility of design as per the requirement. |