Use of cytosolic metabolite patterns to estimate free magnesium in normoxic myocardium

Cytosolic free magnesium (Mg_f) is considered relatively constant. To test this concept, Mg_f was estimated during hyperkalemic ventricular akinesis, normal and maximum adrenergic stimulation, and sulfate loading of the normoxic perfused guinea-pig heart. The Mg_f estimates utilized a new sliding scale derived from the Mg²⁺-dependence of glyceraldehyde-3-phosphate dehydrogenase/phosphoglycerate kinase (GAPDH/PGK). The pseudo constant K_GAPDH′·K_PGK′ was measured as (creatine phosphate]3-phosphoglycerate]lactate]K_LDH/(creatine]P_iglyceraldehyde 3-phosphate]pyruvate]K_CK), which varied with magnesium due to K_CK (CK, LDH = creatine kinase, lactate dehydrogenase). However, the correct magnesium dependencies of the true constants K_GAPDH·K_PGK and K_CK were taken from the literature. The Mg²⁺] at which pseudo K_GAPDH′·K_PGK′ equalled true K_GAPDH·K_PGK was the best estimate of Mg_f. Mg_f fell to ≈0.13 mM in hyperkalemic arrest from a control of ≈0.6 mM, rising to ≈0.85 mM only during maximum adrenergic stress. Mg_f increased further to ≈1.3 mM during sulfate loading which induced ATP catabolism. Mg_f and ATP were reciprocally related. Thus; (1) myocardial free Mg²⁺] judged from GAPDH/PGK mass-action relations changed appreciably only under extreme physiological states; (2) ATP was a major chelator of Mg²⁺ in perfused myocardium, i.e., acute ATP pool size reduction may be associated with increments in Mg_f.