Adenylate kinases of thermophiles and : biochemical and kinetic studies.

Adenylate kinases (AK) play a pivotal role in the regulation of cellular energy. The aim of our work was to achieve the overproduction and purification of AKs from two groups of bacteria and to determine, for the first time, the comprehensive biochemical and kinetic properties of adenylate kinase from Gram-negative (AK) and Gram-positive (AK). Therefore we determined and values, and the effects of temperature, pH, metal ions, donors of the phosphate groups and inhibitor ApA for both thermophilic AKs. The kinetic studies indicate that both AKs exhibit significantly higher affinity for substrates with the pyrophosphate group than for adenosine monophosphate. AK activation by Mg and Mn revealed that both ions are efficient in the synthesis of adenosine diphosphate and adenosine triphosphate; however, Mn ions at 0.2-2.0 mmol/L concentration were more efficient in the activation of the ATP synthesis than Mg ions. Our research demonstrates that zinc ions inhibit the activity of enzymes in both directions, while ApA at a concentration of 10 µmol/L and 50 µmol/L inhibited both enzymes with a different efficiency. Sigmoid-like kinetics were detected at high ATP concentrations not balanced by Mg, suggesting the allosteric effect of ATP for both bacterial AKs.