Electric car-sharing systems have attracted large attention in recent years as a new business model for achieving both economic and environmental benefits in urban areas. Among different types, the one considered in this paper is the so-called one-way car-sharing system whereby a user can begin and end a trip at any station of the system. At the same time, the Vehicle-to-Grid (V2G) concept is emerging as a possible innovative solution for smart power grid control. A management system that combines car-sharing system operations and V2G technology is a recent challenge for academia and industry. In this work, a mixed integer linear programming formulation is proposed to find the optimal management of electric vehicles in a one-way car-sharing system integrated with V2G technology. The proposed mathematical model allows finding the optimal start-of-day electric vehicles distribution that maximizes the total revenue obtained from system users and V2G profits through daily electric vehicles charging/discharging schedules. These schedules are based on mean daily users' electric vehicles requests and electricity prices. The model can be applied to evaluate the possible average daily profitability of V2G operations. In order to test the model performance, we applied it to a small-size test network and a real-size test network (the Delft network in the Netherlands). Under the model assumptions, the adoption of V2G technology allows to fully cover the daily charging costs due to users’ trips and to obtain V2G profits by taking advantage of electric vehicles unused time without significantly reducing the satisfied car-sharing system demand. Most of the energy purchased to charge the electric vehicles batteries is provided back to the grid during energy peak load demand, creating benefits also for energy providers.