Calcium-calmodulin modulation of the activity of Na+/H+ exchanger in human platelets.
This study aimed at establishing the role of calmodulin in regulating pHi of human platelets under acid loads and in stimulated states. The response of human platelets to thrombin was an initial drop of pHi followed by a recovery with a significant increase above the pre-stimulation level in control experiments and a recovery to initial values in platelets maintained in the presence of 19 mmol/l TFP (trifluoperazine = 2 trifluoromethyl-10 [3'-(1 methyl-4-piperazinyl) propyl] phenothiazine). The change in pHi after 8 min was 0.130 +/- 0.030 in the control and 0.001 +/- 0.011 pH units in TFP (P < 0.05). The initial velocity of recovery from an acid load was reduced to 56.7 +/- 6% of the control (n = 6, P < 0.05) with 50 mumol/l W7 (N-(6 aminohexyl)-5-chloro-l-naphthalene sulphonamide), and to 29.7 +/- 4.3% of the control (n = 8, P < 0.05) with 19 mumol/l TFP. The initial velocity of recovery was significantly greater in recalcified platelets than in the preparations kept in the nominal absence of extracellular calcium (1.08 +/- 0.12 vs 0.66 +/- 0.12 pH units per min, P < 0.05). Lower concentration of TFP had an inhibitory effect only in the presence of calcium. The velocities of recovery reached similar values at higher TFP concentration. The significant interaction between Ca2+ and TFP concentrations indicates that the Ca-calmodulin complex, rather than an unspecified direct action of TFP, is responsible for the modulation of the Na+/H+ exchanger. These findings indicate that calcium-calmodulin participates in both the recovery of pH after an acid load and the increase of pHi in stimulated states of human platelets.
Gende
O A
OA
Centro de Investigaciones Cardiovasculares, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Argentina.
D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology; D013997:Time Factors; D014268:Trifluoperazine / Q000494:pharmacology
D002147:Calmodulin; D017923:Sodium-Hydrogen Exchangers; D014268:Trifluoperazine; D000584:Amiloride; D013917:Thrombin; D002118:Calcium; C039614:ethylisopropylamiloride