Enzymatic synthesis of deoxyadenosine triphosphate
Deoxyadenosine triphosphate is able to be enzymatically synthesized with DNA as the starting material using deoxyribonuclease, nuclease P1, adenylate kinase, and pyruvate kinase. The synthesis starts with the heat denaturation of DNA followed by treatment with DNase I to produce oligomers. Next, the solution is treated with nuclease P1 to form deoxynucleoside monophosphates. Using a mixture of adenylate kinase and pyruvate kinase, the deoxyadenosine monophosphate was selectively converted to dATP. After purification, a purity of 90%-95% can be achieved using this method of synthesis with a 40% overall yield.
Health effects
In immunocompromised individuals
High levels of dATP in the body can be toxic and result in impaired immune function, since dATP acts as a noncompetitive inhibitor for the DNA synthesis enzymeribonucleotide reductase. Patients with adenosine deaminase deficiency tend to have elevated intracellular dATP concentrations because adenosine deaminasenormally curbs adenosine levels by converting it into inosine. Deficiency of the enzyme adenosine deaminase is known to cause immunodeficiency in individuals. Research has found that dATP may be a potential toxic metabolite in adenosine deaminase deficiency. Patients in the study who were immunodeficient and adenosine deaminase deficient were found to have over 50 times the levels of dATP in their erythrocytes compared to non-immunodeficient, adenosine deaminase deficient patients. This is abnormal and provides evidence that increased erythrocyte dATP levels are the toxic metabolites responsible for immune system deficiency in individuals with adenosine deaminase deficiency. Infusion of normal, non-enzyme deficient erythrocytes resulted in the loss of dATP in the erythrocytes of these individuals. Cells lacking the ability to transport or phosphorylate dATP have been shown to exhibit increased resistance to the toxic effects of excessive dATP, suggesting that the toxicity of dATP is dependent on the ability to intracellularly phosphorylate dATP. As such, some treatments for ADA focus on reducing dATP phosphorylation by the targeted inhibition of the responsible deoxynucleoside kinases, such as adenosine kinase and deoxycytidine kinase. Deoxycytidine administered intravenously has also been used as an ADA treatment, although a clinical study found that deoxycytidine had only limited clinical effects on the T-cell immunity of ADA patients without discounting that some patients may respond more significantly to deoxycytidine therapy. In cardiac muscle In cardiacmyosin, dATP has been shown to be a viable alternative to ATP as an energy substrate for facilitatingcross-bridge formation. In an experiment involving caninedilated cardiomyopathy, increasing cardiac dATP was found to be a potentially effective treatment for DCM.