Site-Specific Nanocluster Synthesis in Energy-Coupled Biomolecular Hosts
Technical Report,01 Aug 2016,30 Nov 2017
US Army Research Laboratory Aberdeen Proving Ground United States
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Although several proteins have been used to synthesize fluorescent gold metal nanoclusters AuNCs, the fundamental mechanisms of AuNC formation and stabilization in protein conjugates is still a topic of investigation. Understanding key binding residues and protein structures will enable, for the first time, the ability to engineer site-specific nanocluster growth within biomolecular hosts. This report will describe our efforts to elucidate the synthesis and stabilization mechanisms of AuNCs in various protein hosts. Several mutant forms of trypsin were prepared using site-directed mutagenesis, and the mutant proteins purified and their ability to synthesize and stabilize AuNCs will be detailed. In addition, gold-coordinating ligands identified through proteolysis and peptide electrospray ionization-mass spectrometry that may yield insight into the design of novel protein nanoclusters will be explored. Finally, development of 3-D atomic models to predict gold-protein interactions based on experimentally obtained X-ray data will be discussed.
- Genetic Engineering and Molecular Biology
- Atomic and Molecular Physics and Spectroscopy