NDLI: Noninvasive $^{31}P$ NMR probes of free $Mg^{2+},$ MgATP, and MgADP in intact Ehrlich ascites tumor cells

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Noninvasive $^{31}P$ NMR probes of free $Mg^{2+},$ MgATP, and MgADP in intact Ehrlich ascites tumor cells

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Noninvasive $^{31}P$ NMR probes of free $Mg^{2+},$ MgATP, and MgADP in intact Ehrlich ascites tumor cells

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Yushok, W. D. Gupta, R. K.
Abstract	(31)P NMR spectra of Ehrlich ascites tumor cells, suspended in a physiological medium, show well-defined alphaP, betaP, and gammaP resonances of intracellular ATP. A comparison of the separation of 360 +/- 1 Hz between the resonances of alphaP and betaP in intact cells with the corresponding separations of 349 +/- 0.5 and 438 +/- 1 Hz in noncellular MgATP and ATP standards, measured at 10 degrees C and 40.5 MHz NMR frequency, reveals that 88% of the total intracellular ATP (3.3 mM) is complexed to Mg(2+). The corresponding value for intracellular free Mg(2+) is 0.44 mumol/ml out of a total Mg(2+) content of 8.5 mumol/ml cell water. A similar value of free Mg(2+) is obtained from a comparison of the separation of 560 +/- 1 Hz between the betaP and gammaP resonances of intracellular ATP with the corresponding values of 552 +/- 0.5 and 621 +/- 1 Hz in noncellular MgATP and ATP standards. The fraction of total cellular ATP complexed to Mg(2+) decreased to 80% in tumor cells incubated in a medium containing adenosine, glucose, and P(i) without Mg(2+), the corresponding level of intracellular free Mg(2+) being significantly lower (0.24 mM) than that in the unincubated cells. With Mg(2+) present in the incubation medium, MgATP and total Mg(2+) levels increased by approximately 2.5 mM, whereas with no added Mg(2+), the MgATP increased 1.6 mM apparently at the expense of other Mg(2+)-complexed constituents since total Mg(2+) remained essentially unchanged. At the measured levels of free Mg(2+), intracellular ADP will be only 35-50% complexed to Mg(2+). A knowledge of the state of Mg(2+) complexation of ATP and ADP in intact Ehrlich tumor cells allows an accurate estimation of the phosphorylation potential and the extent of deviation of the mass action ratio ([MgATP][AMP](f)/[MgADP][ADP](f)) in cells with high phosphorylation potential from the Mg(2+)-dependent equilibrium value of the adenylate kinase reaction. The significance of the free Mg(2+) value measured by the noninvasive NMR method in relation to the existence of Mg(2+) complexation and compartmentation in tumor cells is discussed.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	5
Volume Number	77
Language	English
Publisher	National Academy of Sciences
Publisher Date	1980-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Adenosine Diphosphate Metabolism Adenosine Triphosphate Carcinoma, Ehrlich Tumor Magnesium Adenosine Animals Cell Compartmentation Cells, Cultured Cytosol Glucose Magnetic Resonance Spectroscopy Mice Phosphates Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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