An ATP "Synthase" Derived from a Single Structural Domain of Bacterial Histidine Kinase

ATP (adenosine triphosphate) is a vital energy source for living organisms, and its biosynthesis and precise concentration regulation often depend on macromolecular machinery composed of protein complexes or complicated multidomain proteins. We have identified a single-domain protein HK853CA derived from bacterial histidine kinases (HK) that can catalyze ATP synthesis efficiently. Here, we explored the reaction mechanism and multiple factors that influence this catalysis through a combination of experimental techniques and molecular simulations. Moreover, we optimized its enzymatic activity and applied it as an ATP replenishment machinery to other ATP-dependent systems. Our results broaden the understanding of ATP biosynthesis and show that the single CA domain can be applied as a new biomolecular catalyst used for ATP supply. A single-domain ATP "synthase" was derived from bacterial histidine kinases. This enzyme catalyzes ATP synthesis from ADP under mild conditions in vitro by exploiting an AdK-like catalytic mechanism. It can be generally applied as new biomolecular catalyst used for ATP supply to facilitate multifarious vital phosphorylation systems. image