A solution of succinic anhydride (SA) in buffer N-tris(hydroxymethyl)methyl-2-aminoethane sulfonic acid (TES), the SATES solution, potentiates the effect of carbonyl containing agonists on frog muscle (del Castillo et al., Br. J. Pharmac. 84: 275-288, 1985). Here we report that the main compound in the SATES solution is a monosuccinyl ester of TES (MST). This compound is not an agonist of the acetylcholine (ACh) receptor nor is it an inhibitor of ACh esterase, yet MST potentiates the ACh-induced tension and depolarization in frog muscle to a new maximum. It increases the amplitude of miniature endplate potentials but has no effect on the time course of miniature endplate currents. The acetylated analogue of MST (mono-acetyl-TES: MAT) had similar but more pronounced effects on frog muscle. Neither MST nor MAT affect [3H]Ach binding to receptor-rich membranes from the electric organ of Torpedo californica, or the affinity state transition induced by agonists. Radiolabeled MST does not bind to these membranes and MAT does not alter agonist-induced ion flux. Therefore, these compounds seem to act as positive modulators of muscle ACh receptor but are inactive in Torpedo vesicles.