In this study, the pre-service chemistry teachers' understanding of potential energy curve was investigated. The subjects were 24 junior students and 26 senior students studying chemistry education in a college of education. A concept questionnaire consisted of thought experiments with different initial conditions was developed to survey the pre-service teachers' conceptions of potential energy curve. The survey results showed that the pre-service chemistry teachers had difficulties to accept the negative values for potential energy and total energy. And they knew the mechanical energy conservation but they could not apply it properly to the thought experiment situations given in the questionnaire. Also they had the knowledge about the direction of force exerted between the two balls, but many of them believed that the balls would stop moving at the bottom of potential energy curve well. In addition, it was discovered that few pre-service teachers could relate the thought experiments to the chemical bonding, the liquefaction of gas, and the molecular vibration.