Glycerol inhibits or uncouples the plasma membrane (Ca(2+)+Mg2+)ATPase of kidney proximal tubules depending on the Ca2+ concentration.
In this report it is shown that glycerol (0.5-10% v/v) stimulate the C12-E9-solubilized renal (Ca(2+)+Mg2+)ATPase in the presence of low concentrations of free Ca2+ (< 10(-6) M). At 4% (v/v), the polyol decreases the K0.5 for Ca2+ from 1.15 to 0.22 microM at the high-affinity site, and a very-high-affinity Ca2+ component appears. This component has a K0.5 < or = 10(-9) M and its maximal velocity is about one-third that of the fully activated enzyme (at 10-20 microM Ca2+), which is not affected by glycerol (21.1 and 20.2 nmollmg-1.min-1 in the absence and presence of the polyol, respectively). The low-affinity, inhibitory component of the Ca2+ curve (50-1000 microM) is also unaffected by glycerol. With 0.07 microM free Ca2+ and soluble enzyme, the stimulatory effect of glycerol saturates at approximately 10% (v/v). In contrast, with 17 microM free Ca2+, glycerol has little effect up to 10% (v/v), and then progressively inhibits ATPase activity. These data indicate that the effect of the polyol is modulated by the occupancy of the high-affinity Ca2+ sites. In native vesicles, the stimulation promoted by low concentrations of glycerol at low concentrations of Ca2+ is accompanied by inhibition of active Ca2+ transport, indicating that, in these conditions, the polyol uncouples ATPase activity and ATP-driven Ca2+ influx.
Sola-Penna
M
M
Departamento de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Brasil.
Lemos
A dos P
Ados P
Vieyra
A
A
D000818:Animals; D001692:Biological Transport; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D005990:Glycerol / Q000494:pharmacology*; D007687:Kidney Tubules, Proximal / Q000201:enzymology*; D007700:Kinetics; D008962:Models, Theoretical; D012756:Sheep
D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D005990:Glycerol; D002118:Calcium