Single molecule studies of enzyme mechanisms

Fluorescence microscopy is an efficient method to study enzyme reactions. In this method, light can be focused in a very small volume to reduce the background of fluorescence with the use of lasers and modern optical techniques. Some of the documented applications of fluorescence microscopy in enzyme reaction study are in flavin enzymes, staphylococcal nuclease, dihydrofolate reductase, conformational heterogeneity, T4 bacteriophage DNA replication, helicases, λ exonuclease and HIV-1 reverse transcriptase. In all these applications, use of fluorescence microscopy made it possible to determine the organization and assembly of complex structures of proteins and nucleic acids without the interference of reactions/interactions occurring in the bulk solution and ensemble averaging.