Previous research indicates that both the extent and timing of muscular activation at the knee can be influenced by muscle activity state, joint angle, weight-bearing status and trunk position. However, little research to date has evaluated protective neuromuscular response times and activation patterns to an imposed perturbation with the knee joint in a functional, weight-bearing stance. Hence, we designed a lower extremity perturbation device to produce a sudden, forward and either internal or external rotation moment of the trunk and femur relative to the weight-bearing tibia. Surface electromyography (EMG) recorded long latency reflex times of the medial and lateral quadriceps, hamstring and gastrocnemius muscles in 64 intercollegiate lacrosse and soccer players in response to both internal and external rotation perturbation. We found the gastrocnemius fired significantly faster that the hamstring, which in turn fired significantly faster than the quadriceps. There was also a significant difference in activation times of the medial and lateral hamstring not found for the quadriceps or gastrocnemius muscles. Our findings confirmed that reactive neuromuscular responses following this functional perturbation differ markedly from those previously reported using seated, partial weight-bearing and/or uni-planar models under relaxed conditions.