Mitochondrial adenosine triphosphatase from human placenta--inhibition by free magnesium ions of ITP hydrolysis.
The effects of Mg2+ and bicarbonate on the kinetics of ITP hydrolysis by soluble ATPase (F1) from human placental mitochondria were studied. Increasing amounts of Mg2+ at fixed ITP concentration, caused a marked activation of F1 followed by inhibition at higher Mg2+ concentration. The appropriate substrate for the mitochondrial F1 seems to be the MgITP complex as almost no ITP was hydrolysed in the absence of magnesium. Mg2+ behaved as a competitive inhibitor towards the MgITP complex. In this respect the human placental enzyme differ from that from other sources such as yeast, beef liver or rat liver. The linearity of the plot presenting competitive inhibition by free Mg2+ of MgITP hydrolysis (in the presence of activating bicarbonate anion) suggests that both Mg2+ and MgITP bind to the same catalytic site (Km(MgITP) = 0.46 mM, Ki(Mg) = 4 mM). When bicarbonate was absent in the ITPase assay, placental F1 exhibited apparent negative cooperativity in the presence of 5 mM Mg2+, just as it did with MgATP as a substrate under similar conditions. Bicarbonate ions eliminated the negative cooperativity with respect to ITP (as the Hill coefficient of 0.46 was brought to approx. 1), and thus limited inhibition by free Mg2+. The results presented suggest that the concentration of free magnesium ions may be an important regulatory factor of the human placental F1 activity.
Department of Biochemistry Medical Academic School, Gdańsk, Poland.
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Aleksandrowicz
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D001639:Bicarbonates / Q000494:pharmacology; D001665:Binding Sites / Q000502:physiology; D002384:Catalysis; D005260:Female; D006801:Humans; D006868:Hydrolysis; D007293:Inosine Triphosphate / Q000378:metabolism*; D016014:Linear Models; D008274:Magnesium / Q000494:pharmacology*; D008928:Mitochondria / Q000201:enzymology*; D010920:Placenta / Q000201:enzymology* / Q000648:ultrastructure; D011247:Pregnancy
D001639:Bicarbonates; D007293:Inosine Triphosphate; D000251:Adenosine Triphosphatases; D008274:Magnesium