On February 28, the research team of Li Conggang and Yang Minghui from the Institute of Precision Measurement Science and Technology Innovation, Chinese Academy of Sciences, made progress in the development and application of ATP synthetase, obtained the ATP synthetase with a single domain for the first time, analyzed the molecular mechanism of enzyme catalysis, and applied it to the phosphorylation experiment of various substrate molecules.
ATP is the main source of energy in living organisms and is crucial for life activities. The synthesis of ATP in living organisms relies on the ATP synthase protein machine and the multi domain adenosine kinase system, but the system is large and difficult to scale up. The commonly used microbial fermentation and enzyme catalysis methods in industrial production have drawbacks such as cumbersome processes, complex operations, and difficult control. Therefore, the development of efficient ATP synthase with simple and mild reaction conditions is of great significance for achieving recoverable energy supply of ATP in vitro and biological systems.
The histidine kinase HK853 from marine thermophilic fungi is a protease with multiple catalytic functions, exhibiting good thermal stability and substrate specificity. Research has found that the CA domain of HK853 has a new ATP synthesis function, which can efficiently synthesize ATP using ADP, and the catalytic reaction is mild and controllable. Based on this, researchers designed the single domain ATP synthase HK853CA and characterized the optimal conditions for HK853CA to achieve ATP synthesis catalytic reaction using techniques such as nuclear magnetic resonance, such as temperature, concentration, pH value, and metal ion binding ability. The study combined experiments and molecular dynamics simulations to elucidate the molecular mechanism of enzyme catalysis, namely that HK853CA can simultaneously bind two molecules of ADP to generate ATP and AMP, and the reaction is reversible under certain conditions. The study applied the ATP synthase HK853CA to a biological system for phosphorylation of various substrates, such as proteins, DNA, and small molecules, achieving efficient utilization of ADP and ATP energy molecules.
This study designed a series of mutations based on the molecular mechanism of enzyme catalysis to regulate its catalytic activity, and found that this catalytic reaction has a certain universality in the histidine kinase family, providing a new research direction for subsequent enzyme modification and structural optimization.
The related research results, titled An ATP "Synthesis" Derived from a Single Structural Domain of Bacterial Histidine Kinase, were published in the German Journal of Applied Chemistry. The research work was supported by the National Natural Science Foundation of China and the Chinese Academy of Sciences strategic leading science and technology project (Category B).
Single domain ATP synthase designed based on bacterial histidine kinase
