Chronic ankle instability (CAI) is not only an ankle issue, but also affects sensorimotor system. People with CAI show altered muscle activation in proximal joints such as hip and knee. However, evidence is limited as controversial results have been presented regarding changes in activation of hip muscles in CAI population. PURPOSE: To investigate the effect of CAI on activity of hip muscles during normal walking and walking with perturbations. METHODS: 8 subjects with CAI (23 ± 2 years, 171 ± 7 cm and 65 ± 4 kg) and 8 controls (CON) matched by age, height, weight and dominant leg (25 ± 3 years, 172 ± 7 cm and 65 ± 6 kg) walked shoed on a split-belt treadmill (1 m/s). Subjects performed 5 minutes of baseline walking and 6 minutes walking with 10 perturbations (at 200 ms after heel contact with 42 m/s² deceleration impulse) on each side. Electromyography signals from gluteus medius (Gmed) and gluteus maximus (Gmax) were recorded while walking. Muscle amplitudes (Root Mean Square normalized to maximum voluntary isometric contraction) were calculated at 200 ms before heel contact (Pre200), 100 ms after heel contact (Post100) during normal walking and 200 ms after perturbations (Pert200). Differences between groups were examined using Mann Whitney U test and Bonferroni correction to account for multiple testing (adjust α level p≤ 0.0125). RESULTS: In Gmed, CAI group showed lower muscle amplitude than CON group after heel contact (Post100: 18±7 % and 47±21 %, p< .01) and after walking perturbations ( 31±13 % and 62±26 %, p< .01), but not before heel contact (Pre200: 5±2 % and 11±10 %, p= 0.195). In Gmax, no difference was found between CAI and CON groups in all three time points (Pre200: 12±5 % and 17±12 %, p= 0.574; Post100: 41±21 % and 41±13 %, p= 1.00; Pert200: 79±46 % and 62±35 %, p= 0.505). CONCLUSIONS: People with CAI activated Gmed less than healthy control in feedback mechanism (after heel contact and walking with perturbations), but not in feedforward mechanism (before heel contact). Less activation on Gmed may affect the balance in frontal plane and increase the risk of recurrent ankle sprain, giving way or feeling ankle instability in patients with CAI during walking. Future studies should investigate the effect of Gmed strengthening or neuromuscular training on CAI rehabilitation.