Measurement of the dissociation constant of MgATP at physiological nucleotide levels by a combination of 31P NMR and optical absorbance spectroscopy.

Abstract:

:To resolve a controversy in the literature concerning the affinity of Mg++ for ATP to be used in our noninvasive 31P NMR procedure for the determination of free Mg++ in living cells, we have reinvestigated the apparent dissociation constant of MgATP under physiologic ionic conditions and over the cellular range of ATP concentrations by a combination of NMR and optical absorbance techniques. The new combination method utilizes 31P NMR chemical shifts to determine the degree of Mg++ chelation of ATP in a solution containing free ATP and MgATP, and uses a properly calibrated indicator dye, antipyrylazo III, for optical measurement of free Mg++ in the same solution. The data yield an average value of 50 +/- 10 microM for the apparent dissociation constant of MgATP which indicates low levels of free Mg++ (less than 1 mM) in several different types of tissues, including perfused heart muscle, contrary to a recent report in the literature.

authors

Gupta RK,Gupta P,Yushok WD,Rose ZB

doi

10.1016/0006-291x(83)91562-0

subject

Has Abstract

pub_date

1983-11-30 00:00:00

pages

210-6

issue

1

eissn

0006-291X

issn

1090-2104

pii

0006-291X(83)91562-0

journal_volume

117

pub_type

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