Three-step preparation and purification of phosphorus-33-labeled creatine phosphate of high specific activity.
Rabbit heart mitochondria were used as a source of enzymes for the synthesis of phosphorus-labeled creatine phosphate. This method is based on the coupled reaction between mitochondrial oxidative phosphorylation and mitochondrial-bound creatine kinase. It is possible to convert more than 90% of the inorganic phosphate (Pi) to creatine phosphate. The method used only small amounts of adenine nucleotides which led to a product with only slight nucleotide contamination. This could be removed by activated charcoal extraction. For further purification, a method for the removal of residual Pi is described.
D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D066298:In Vitro Techniques; D007553:Isotope Labeling / Q000379:methods*; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D010725:Phosphocreatine / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D011485:Protein Binding; D011817:Rabbits
D010761:Phosphorus Radioisotopes; D010725:Phosphocreatine; D003402:Creatine Kinase