Stimulation of the Ca(2+)-ATPase of sarcoplasmic reticulum by disulfiram.
Disulfiram [bis(diethylthiocarbamoyl)disulphide] has been found to stimulate reversibly the Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum. At pH 7.2, 2.1 mM ATP and 25 degrees C, ATPase activity was found to double on addition of 120 microM disulfiram. Stimulation fitted to binding of disulfiram at a single site with a Kd of 61 microM. Disulfiram had no effect on the Ca2+ affinity of the ATPase or on the rate of phosphorylation of the ATPase by ATP, but increased the rate of dissociation of Ca2+ from the phosphorylated ATPase (the transport step) and increased the rate of dephosphorylation of the phosphorylated ATPase. It also decreased the level of phosphorylation of the ATPase by Pi, consistent with a 7.5-fold decrease in the equilibrium constant of the phosphorylated to non-phosphorylated forms (E2PMg/E2PiMg) at 80 microM disulfiram. Disulfiram had no significant effect on the concentration of ATP resulting in stimulation of ATPase activity, suggesting that it does not bind to the empty nucleotide-binding site on the phosphorylated ATPase. Studies of the effects of mixtures of disulfiram and jasmone (another molecule that stimulates the ATPase) suggest that they bind to separate sites on the ATPase.
Starling
A P
AP
Department of Biochemistry, University of Southampton, U.K.
East
J M
JM
Lee
A G
AG
D000427:Alcohol Deterrents / Q000494:pharmacology*; D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D004221:Disulfiram / Q000494:pharmacology*; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D018482:Muscle, Skeletal / Q000201:enzymology; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000187:drug effects* / Q000201:enzymology
D000427:Alcohol Deterrents; D010710:Phosphates; D000252:Calcium-Transporting ATPases; D004221:Disulfiram