Identification of ATP diphosphohydrolase activity in human term placenta using a novel assay for AMP.
Human term placenta contains an ATP diphosphohydrolase activity which hydrolyses ATP to ADP and inorganic phosphate and ADP to AMP and a second mole of inorganic phosphate. The activity has a pH optimum between 8.0 and 8.5. Magnesium or calcium ions are required for maximum activity. Other nucleoside phosphates, p-nitrophenyl phosphate or sodium pyrophosphate, are not hydrolysed. The activity is not due to ATPases, or to myokinase, as determined by the use of inhibitors. NaF and NaN3 were found to inhibit strongly the activity thus identifying it as an ATP diphosphohydrolase. A sensitive enzymatic assay for measurement of AMP, one of the products of the reaction, was established, based on the strong inhibition of muscle fructose 1,6-biphosphatase by AMP. The range of the assay was 0.05-0.8 microM AMP. ATP diphosphohydrolase was found to have a rate of AMP production from ADP twice the rate from ATP. Under the same conditions, the assay for Pi release, on the other hand, gave velocities similar to each other for the two substrates. The activity appears to be identical to the ADP-hydrolysing activity in placenta reported by others.
Papamarcaki
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Laboratory of Biological Chemistry, University of Ioannina Medical School, Greece.
Tsolas
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O
D000249:Adenosine Monophosphate / Q000378:metabolism*; D001081:Apyrase / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D006597:Fructose-Bisphosphatase / Q000037:antagonists & inhibitors; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008274:Magnesium / Q000494:pharmacology; D010920:Placenta / Q000187:drug effects / Q000201:enzymology*; D011188:Potassium / Q000494:pharmacology; D013379:Substrate Specificity
D000249:Adenosine Monophosphate; D006597:Fructose-Bisphosphatase; D001081:Apyrase; D008274:Magnesium; D011188:Potassium; D002118:Calcium