Stepwise conformational transition of crystalline disodium adenosine 5′-triphosphate with relative humidity as studied by high resolution solid state 13C and 31P NMR

Abstract

Stepwise conformational transition of disodium adenosine 5′-triphosphate (Na 2ATP) crystals as a function of relative humidity (RH), was examined by means of high resolution solid state NMR spectroscopy. The presence of two types of molecules, A and B, with altered conformations in an asymmetric unit, is evident from both 31P and 13C NMR spectra, irrespective of the three different crystalline forms, mono-, di- and trihydrates, in which, cell parameters changed linearly with RH. Hydration-dependent conformational transition from the monohydrate (RH 0–10%) through dihydrate (20–40%) to trihydrate (60–90%), was well monitored by stepwise upfield displacements of the C1′ 13C signals of ribose moiety (molecule B), although, the corresponding peak for the molecule A, is almost unchanged. This means that, adaptation of stepwise structural transitions to the linear expansion of cell parameters is almost entirely ascribed to the conformational readjustment of the molecule B, together with varying proportion of two types of hydrate forms, at the RH range of 20–40%. We consider that, small clusters or thin layers are formed in the beginning of the transition and subsequently, the number of them, rather than the size or thickness increases, because the phosphorous spin–lattice relaxation times, T 1 P, for the three phosphorous nuclei, including those of mixed state, of the two hydrate forms, turned out to be similar due to sufficient fast spin diffusion rates among them. Since only one direction of the cell parameters is changed, as determined by the previous X-ray diffraction study, we conclude that thin layer type micro crystals might be produced during the stepwise transition. Further, it was found that, these molecules acquired motional flexibility mainly at the bound water molecules, together with increased relative humidity, as determined by a variety of relaxation parameters.