Anionic (NO(3) (-), Br(-), I(-), and SCN(-)) and cationic (Zn(++) and Cd(++)) potentiators of the twitch output of skeletal muscle depress the active binding of Ca by sarcoplasmic reticulum isolated from rabbit skeletal muscle. Zinc and Cd exchange for Ca and Mg at the binding sites of the reticular membranes, whereas the anions effectively induce a replacement by Mg of Ca bound actively in the presence of ATP. In the absence of ATP, the passive binding of both Ca and Mg is increased by the anions tested. Furthermore, the anions increase the total capacity of the membrane fragments for passive cation binding. The Ca-stimulated ATPase activity of the membranes is inhibited by Zn and Cd, but not by the anions. Shifts in cations bound to muscle membrane systems caused by agents that increase the force of contraction developed during the twitch are considered to be the primary event modifying excitation-contraction coupling, and thus leading to potentiation.