Machining-induced surface integrity has an important effect on reliability and service life of the components used in the aerospace industry where titanium alloy Ti-6Al-4V is widely applied. Characterisation of machining-induced surface integrity and revealing its effect on fatigue life are conducive to structural fatigue life optimisation design. In the present study, surface topography, residual stress, microstructure and micro-hardness were first characterised in peripheral milling of titanium alloy Ti-6Al-4V. Then, low-cycle fatigue performances of machined specimens were investigated on the basis of the tension-tension tests. Finally, the effects of surface integrity factors (stress concentration factor, residual stress and micro-hardness) on fatigue performances were discussed. Results show that stress concentration can reduce the fatigue life while increasing the residual compressive stress, and micro-hardness is beneficial to prolonging the fatigue life, but when the surface material of the specimen is subjected to plastic deformation due to yield, the residual stress on the surface is relaxed, and the effect on the fatigue performance is disappeared. Under the condition of residual stress relaxation, the stress concentration factor is the main factor to determine the low-cycle fatigue life of titanium alloy Ti-6Al-4V. While for the specimens with no residual stress relaxation, micro-hardness was the key factor to affect the fatigue life, followed by residual stress and stress concentration factor, respectively.