Experimental 31P NMR study of the influence of ionic strength on the apparent dissociation constant of MgATP

Mottet, I.;Demeure, R.;Gallez, Bernard;Grandin, Cécile;Pringot, J.;et.al.
(1994) Magnetic Resonance Materials in Physics, Biology and Medicine — Vol. 2, n° 2, p. 101-107 (1994)

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Authors
  • Mottet, I.UCLouvain
    Author
  • Demeure, R.UCLouvain
    Author
  • Author
  • Grandin, CécileUCLouvain
    Author
  • Van Beers, BernardUCLouvain
    Author
  • Pringot, J.UCLouvain
    Author
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Abstract
The classical method for31P NMR determination of intracellular free magnesium concentration ([Mg 2+free]) requires an accurate knowledge of the apparent dissociation constant (K D ) of MgATP. There is a large difference between the previously determined values ofK D . Although the value of 50 µM, determined by a31P NMR method, is now largely accepted, a value of 86 µM has more recently been measured with a fitting method derived from the original one, and with a different ionic strength. The purpose of our study was to assess if the cause of the difference between these two previously reportedK D values was due to the measuring method or to the ionic strength value used. Working at pH=7.2,T=37°C, and [KCl]=0.25 M, we performedK D measurements with the original31P NMR method and with the fitting method. The results (67±13 µM and 61±20 µM, respectively) were not significantly different. Then, with the first method, we measured KD at [KCl]=0.12 M and found a value of 19±5 µM. We conclude that the main cause of difference between theK D values measured by31P NMR reside in the disparity of ionic strength values used for their measurement. OurK D measurements at [KCl]=0.25 and 0.12 M demonstrate the importance of the ionic strength value used for imitating the intracellular medium on the absolute value of ([Mg 2+free]) measured by31P NMR spectroscopy.
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Citations

Mottet, I., Demeure, R., Gallez, B., Grandin, C., Van Beers, B., & Pringot, J. (1994). Experimental 31P NMR study of the influence of ionic strength on the apparent dissociation constant of MgATP. Magnetic Resonance Materials in Physics, Biology and Medicine, 2(2), 101-107. https://doi.org/10.1007/bf01753065 (Original work published 1994)