Similarities between the effects of dimethyl sulfoxide and calmodulin on the red blood cell Ca2(+)-ATPase.
The Ca2(+)-ATPase of the erythrocyte plasma membrane can be activated by calmodulin, acidic phospholipids, limited proteolysis and self-association. Recently, it has been shown that different organic solvents increase both the Vmax and the Ca2+ affinity of the enzyme (Benaim, G. and De Meis, L. (1989) FEBS Lett. 244, 484-486). In this report the effects of calmodulin and dimethyl sulfoxide (20%, v/v) on the Ca2(+)-ATPase are compared. Dimethyl sulfoxide also elicits the appearance of the low-affinity binding site, which in this enzyme is strictly dependent on calmodulin. Dimethyl sulfoxide increases the Ca2+ affinity of the enzyme in a manner similar to that observed with the use of calmodulin and of acidic phospholipids. This was tested using both native and partially trypsinized ATPase. When activated by calmodulin the enzyme is inhibited by compound 48/80, trifluoperazine and calmidazolium. When activated by dimethyl sulfoxide the enzyme is still inhibited by calmidazolium but is no longer inhibited by either compound 48/80 or trifluoperazine. Activation of the ATPase promoted by either calmodulin or dimethyl sulfoxide is abolished when the Ca2+ concentration is raised from 10 microM to 2 mM. The effect of dimethyl sulfoxide is also abolished by 20 mM Pi. In the presence of 1 to 10 mM Ca2+ the ATPase catalyzes an ATP in equilibrium Pi exchange. The rate of exchange increases several fold when dimethyl sulfoxide is included in the assay medium.
Benaim
G
G
Centro de Biologia Celular, Faculdad de Ciencias, Universidad Central de Venezuela, Caracas.
de Meis
L
L
D000255:Adenosine Triphosphate / Q000097:blood; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000097:blood*; D002147:Calmodulin / Q000494:pharmacology*; D002417:Cattle; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D010710:Phosphates / Q000097:blood / Q000494:pharmacology; D014268:Trifluoperazine / Q000494:pharmacology; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
D002147:Calmodulin; D007093:Imidazoles; D010710:Phosphates; D014268:Trifluoperazine; D003189:p-Methoxy-N-methylphenethylamine; C031938:calmidazolium; D000255:Adenosine Triphosphate; D000252:Calcium-Transporting ATPases; D002118:Calcium; D004121:Dimethyl Sulfoxide