Calcium entry across the microvillous membrane of the human placental syncytiotrophoblast is the first step in the transfer of this important nutrient to the fetus. Calcium uptake by isolated microvillous membranes was time dependent. Equilibrium uptake was very much greater than could be explained by equilibration of the vesicle space with medium, indicating that calcium is bound to internal sites. Addition of the ionophore A23187 greatly increased the rates of influx and efflux, indicating that transport across the plasma membrane is rate limiting in entry. Concentration dependence data for calcium transport at 4 s fit well to a Michaelis-Menten equation having two saturable sites and diffusion. Calcium entry by both transporters was unaffected by calcium channel blockers but was strongly inhibited by the group II metals. The distinct inhibition constant values for strontium inhibition provided additional evidence for two transporters. Calcium binding fit well to a single-site model saturable in the micromolar range. In vivo, the saturable transport processes may mediate calcium entry into the syncytiotrophoblast and binding may regulate concentration within the placental microvilli.