The non-covalent interaction of protein-protein interaction dynamics has been the core research focus of cell signaling and protein functionality.  The non-covalent interaction is more dynamic rather than static, and the protein-protein interaction and recognition complex involve constant conformational fluctuations that regulating the fluctuating interactions at the intra-domain, inter-domain, interface, and overall protein complexes, beyond the conventional on-off binding interaction rate constants.  We have explored the protein-protein interaction energy landscape fluctuations and entanglements before, during, and after the protein-protein recognition complex formed.  We have explored the importance of the energy landscape entanglement and its fluctuations for protein-protein complex function and activities, using our home-developed AFM/Magnetic Tweezers force manipulation combined single-molecule fluorescence imaging nanoscopy.  We specifically studied the mechanisms and dynamics of enzymatic reactions involved with kinase, nitric oxide, and calmodulin proteins, observing the protein conformational fluctuations and the associated structural changes of individual protein molecules under pico-Newton force manipulations simultaneously in real-time by single-molecule FRET detections.  We have also observed significant changes of the enzymatic energy landscape and reaction pathways, while there is a significant tolerance of the enzyme activity to the enzyme conformational perturbation, interrogating direct influence of a mechanical picoNewton(pN) oscillating force on enzymatic activity and the conformational dynamics of the single enzyme under different oscillation frequencies and under enzymatic conditions. Our new multimodal chemical imaging and manipulation macroscopy have a high and broad impact on applications in studying protein structure-dynamics-function relationship.<br/> <br/> <br/>

Nanoscale-manipulation of the force field fluctuation and energy landscape entanglement in protein non-covalent recognition dynamics

2019 Fall National Meeting

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