The contraction of smooth muscle is influenced by the substrate MgATP and the product MgADP. The effects on force, shortening velocity and ATP-turnover, are consistent with an influence on the kinetics of cross-bridge cycling. Part of these effects are mediated via an influence on the regulation of contraction by myosin light chain phosphorylation. Results from preparations activated by thiophosphorylation, show that MgATP and MgADP also interact directly at the cross-bridge level, and are consistent with MgADP acting as a competitive ATP-analogue. The slow shortening velocity and decreased rate of ATP-induced relaxation from rigor in the presence of MgADP, suggest an inhibition of cross-bridge detachment. The rate of ATP-turnover was decreased in the presence of the nonhydrolyzable ATP-analogue AMPPNP. These results may contribute to the characterization of the biochemical reactions in the structurally organized smooth muscle contractile system. In addition, the influence of MgATP and MgADP on smooth muscle contraction suggest that the concentrations of substrate and products, at the level of the contractile proteins, may constitute important regulatory factors in vivo under conditions, such as hypoxia and ischemia, associated with impaired cellular energy supply.