Abstract
Recently we demonstrated that trapping single proteins from solution in a plasmonic double nanohole (DNH) and monitoring the intensity changes of the transmitted light makes it possible to resolve their conformational changes. The power of this technique lies in the possibility of studying unmodified enzymes at work for several minutes up to hours with a temporal resolution of at least 40 μs. Here, we show that for active enzymes that undergo conformational changes, a combination of automated signal analysis and kinetic modelling allows to take a step further: reconstructing the reaction cycle from the detected sequence of conformations.Two llustrative examples will be discussed: adenylate kinase (AdK) and citrate synthase (CS).
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