Observation of Mg2+.ATP and uncomplexed ATP in slow exchange by 31P-NMR at high magnetic fields.

ABSTRACT

The 31P-NMR lines of the beta-phosphate groups in Mg2+.ATP and in metalfree ATP can be observed separately up to 280 K at 8.5 T and up to 285 K at 11.7 T. At 274 K and 8.5 T the beta-phosphorous resonances are in slow exchange at pH values above pH 5, the gamma-phosphorous resonances are in slow exchange only near pH 6, but in fast exchange at low and high pH-values. The fast exchange condition holds for the alpha-phosphorous resonances over the entire pH-range. For Ca2+.ATP and metalfree ATP always fast exchange prevails down to the freezing point of water even at 11.7 T. Based on the separate observation of the 31P-NMR signals of Mg2+.ATP and uncomplexed ATP new experiments are proposed and possible sources of error in 'in vivo' NMR studies are discussed.