... subunits at the phosphorylation step.AbbreviationsAMPPCP, adenylyl-(b,c-methylene)-diphosphonate; E1 P, high-energy phosphoenzyme; E2 P, low-energy phosphoenzyme; EP,phosphoenzyme; K0.5, ... 0.5 mM pNPP, 0.3, 1 or3mM22NaCl, 0.1 mM RbCl, 1% ethanol, with or without 10 lMoligomycin. After the addition of pNPP, the mixture was centri-fuged. The detailed procedure is described ... aliquid scintillation spectrophotometer. The amount of EPwas calculated from the difference between radioactivities ofreaction mixtures with native and acid-denatured enzymes.Determination of...