Probing Enzyme-Surface Interactions via Protein Engineering and Single-Molecule Techniques
Technical Report,01 Apr 2012,31 Mar 2017
University of Colorado - Boulder Boulder United States
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The overall objective of this research is to exploit protein engineering and fluorescence single molecule methods to enhance our understanding of the interaction of proteins and surfaces. Given this objective, the specific aims of this research are to 1 exploit the incorporation of unnatural amino acids in proteins to introduce single-molecule probes i.e., fluorophores for fluorescence resonance energy transfer FRET measurements, 2 demonstrate the utility of site-specific labeling for characterizing surface-induced conformational-bias of proteins at the single-molecule level, and 3 correlate changes in protein structure and interfacial dynamics i.e., adsorption, diffusion, and desorption upon interaction of freely diffusing proteins with surfaces with polar, non-polar, and polymer chemistries.