The mechanical properties of Ti-6Al-4V alloy are sensitive to strain rate and temperature load. The finite element simulation results of high-speed machining Ti-6Al-4V alloy depend on the accurate description of dynamic deformation. However, it is hard to describe the flow stress behavior in current constitutive models in a complex high-speed machining process for Ti-6Al-4V alloy. In this paper, the stress-strain curves of Ti-6Al-4V alloy under the wide ranges of strain rate and temperature are obtained by high-velocity uniaxial impact tests. The apparent coupling between temperature and strain is observed, which proves that the temperature is dependent on a hardening effect for Ti-6Al-4V alloy. A function describing the coupling between temperature and strain is then introduced into the modification for the original Johnson-Cook (JC) constitutive model. The maximum deviation between the predicted data from using the proposed modified JC constitutive model and experimental data is reduced from 10.43% to 4.19%. It can be concluded that the modified JC constitutive model is more suitable to describe the temperature-dependent hardening effect, which provides strong support for accurate finite element simulation of high-speed machining Ti-6Al-4V alloy.