The phosphatase activity of the plasma membrane Ca2+ pump. Activation by acidic lipids in the absence of Ca2+ increases the apparent affinity for Mg2+.
The purified PMCA supplemented with phosphatidylcholine was able to hydrolyze pNPP in a reaction media containing only Mg(2+) and K(+). Micromolar concentrations of Ca(2+) inhibited about 75% of the pNPPase activity while the inhibition of the remainder 25% required higher Ca(2+) concentrations. Acidic lipids increased 5-10 fold the pNPPase activity either in the presence or in the absence of Ca(2+). The activation by acidic lipids took place without a significant change in the apparent affinities for pNPP or K(+) but the apparent affinity of the enzyme for Mg(2+) increased about 10 fold. Thus, the stimulation of the pNPPase activity of the PMCA by acidic lipids was maximal at low concentrations of Mg(2+). Although with differing apparent affinities vanadate, phosphate, ATP and ADP were all inhibitors of the pNPPase activity and their effects were not significantly affected by acidic lipids. These results indicate that (a) the phosphatase function of the PMCA is optimal when the enzyme is in its activated Ca(2+) free conformation (E2) and (b) the PMCA can be activated by acidic lipids in the absence of Ca(2+) and the activation improves the interaction of the enzyme with Mg(2+).
Mazzitelli
Luciana R
LR
IQUIFIB-Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, 1113 Buenos Aires, Argentina.
Adamo
Hugo P
HP
D009597:4-Nitrophenylphosphatase / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001923:Brain Chemistry; D002118:Calcium / Q000502:physiology*; D000252:Calcium-Transporting ATPases / Q000097:blood / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology*; D004789:Enzyme Activation; D008055:Lipids / Q000494:pharmacology*; D008274:Magnesium / Q000378:metabolism*; D010710:Phosphates / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D013552:Swine; D014638:Vanadates / Q000494:pharmacology
D008055:Lipids; D010710:Phosphates; D010713:Phosphatidylcholines; D014638:Vanadates; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009597:4-Nitrophenylphosphatase; D000252:Calcium-Transporting ATPases; D008274:Magnesium; D011188:Potassium; D002118:Calcium