Dependence on intracellular pH and Mg2+ of metabolic compartmentation of phosphoinositides in human erythrocytes.
Effects of intracellular pH and Mg2+ on turnover and extent of metabolic compartmentation of phosphomonoester groups of phosphoinositides and phosphatidate were investigated in human erythrocytes by short-term and equilibrium labeling with [32P]Pi under steady-state conditions. At pH 6.7, the specific radio-activities of phosphoinositides reached apparent equilibrium values, in the range of 70% of that ATP-gamma-P after long-term labelling. At pH 7.2, these values were in the range of 40-50% of that ATP-gamma-P. This demonstrates a decreased accessibility of phosphoinositides to enzymatic phosphorylation and dephosphorylation at the transition from acidic to normal incubation conditions. These changes were more pronounced at pH 7.8. High intracellular [Mg2+] initially activated the turnover of phosphoinositides at pH 7.2. The activation changed into an inhibition and an increase of metabolic compartmentation after three hours preincubation of erythrocytes at high [Mg2+]. The long-term Mg2+ effects are reversible to a great extent by a subsequent re-reduction of [Mg2+]. In conclusion, deprotonation of phosphoinositides by low [H+] or high intracellular [Mg2+] at normal pH induces a decreased accessibility for their specific lipid kinases and phosphatases. This effect may be the result of lateral phase separation of acidic phospholipids as a consequence of divalent cation complexation under both experimental conditions, high pH and high [Mg2+] at normal pH.
Müller
E
E
Institute of Biochemistry II, Medical Faculty, Friedrich Schiller University, Jena, Germany.
Hegewald
H
H
Klinger
R
R
Wetzker
R
R
Cumme
G A
GA
Frunder
H
H
D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism*; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D010766:Phosphorylation
D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols; D008274:Magnesium