Sun, L. et al. J. Am. Chem. Soc. 139, 17525–17532 (2017).

Enzyme dynamics and function are directly correlated in a variety of biological processes, but imaging the dynamics of a high number of single enzymes in real time is challenging. Sun et al. translated 3D movements that are difficult to track with TIRF microscopy into 2D movements on a supported lipid bilayer that is fixed on a coverslip. The researchers used cholesterol-bound DNA that integrates into this bilayer and forms stationary cholesterol–DNA origami rafts. One of the enzymes of interest is statically bound to this DNA origami raft, while the second enzyme of interest is tethered to individual cholesterol molecules and therefore remains freely diffusive within the membrane. This enables enzyme colocalization detection via FRET analysis. The researchers applied this tool to visualize the glucose oxidase–catalase cascade.