ATP formation from adenyl-5'-yl imidodiphosphate, a nonhydrolyzable ATP analog.
The purity of several preparations of adenyl-5'-yl imidodiphosphate (AMP-PNP) was analyzed using thin layer chromatography and the luciferin-luciferase assay. Three contaminants were identified: adenyl-5'-yl phosphoramidate, phosphorylated AMP-PNP, and ATP. The level of ATP contamination ranged from 0.02% to 0.3% in commercially obtained AMP-PNP preparations, and rose to 10% following incubation of AMP-PNP at 37 degrees C for 3 weeks in aqueous solution. The chemistry of the phosphoramidate bond is reviewed briefly, and evidence for a simple mechanism for the spontaneous formation of ATP from AMP-PNP is presented.
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000032:analysis*; D000266:Adenylyl Imidodiphosphate / Q000032:analysis*; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002855:Chromatography, Thin Layer; D001517:Coleoptera / Q000201:enzymology; D007700:Kinetics; D008156:Luciferases
D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D008156:Luciferases