Artificial cells with all-aqueous droplet-in-droplet structures for spatially separated transcription and translation.

The design of functional artificial cells involves compartmentalizing biochemical processes to mimic cellular organization. To emulate the complex chemical systems in biological cells, it is necessary to incorporate an increasing number of cellular functions into single compartments. Artificial organelles that spatially segregate reactions inside artificial cells will be beneficial in this context by rectifying biochemical pathways.

In vitro one-pot construction of influenza viral genomes for virus particle synthesis based on reverse genetics system.

The reverse genetics system, which allows the generation of influenza viruses from plasmids encoding viral genome, is a powerful tool for basic research on viral infection mechanisms and application research such as vaccine development. However, conventional plasmid construction using Escherichia coli (E.coli) cloning is time-consuming and has difficulties handling DNA encoding genes toxic for E.

Engineering of IF -susceptive bacterial F -ATPase.

IF , an inhibitor protein of mitochondrial ATP synthase, suppresses ATP hydrolytic activity of F . One of the unique features of IF is the selective inhibition in mitochondrial F (MF ); it inhibits catalysis of MF but does not affect F with bacterial origin despite high sequence homology between MF and bacterial F . Here, we aimed to engineer thermophilic Bacillus F (TF ) to confer the susceptibility to IF for elucidating the molecular mechanism of selective inhibition of IF .

The series of conformational states adopted by rotorless F-ATPase during its hydrolysis cycle.

FF ATP synthase interchanges phosphate transfer energy and proton motive force via a rotary catalytic mechanism and isolated F-ATPase subcomplexes can also hydrolyze ATP to generate rotation of their central γ rotor subunit. As ATP is hydrolyzed, the F-ATPase cycles through a series of conformational states that mediates unidirectional rotation of the rotor. However, even in the absence of a rotor, the α and β subunits are still able to pass through a series of conformations, akin to those that generate rotation.