Understanding how enzymes function requires a thorough characterization of enzymatic dynamics. Traditional enzymatic assays average over an ensemble of molecules, making it difficult to detect reaction intermediates and conformational fluctuations of the enzyme. These problems can be overcome by observing enzymes functioning in real time on the single-molecule level. This chapter describes recent research efforts to measure singlemolecule enzyme kinetics and observe the structural dynamics of enzymes and discusses new approaches to study multiprotein complexes on the single-molecule level.
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