This study aimed to quantify the influence of an increase in power output (PO) on joint kinematics and electromyographic (EMG) activity during an incremental test to exhaustion for a population of professional cyclists. The hip flexion/extension and internal/external rotation as well as knee abduction/adduction ranges of motion were significantly decreased at 100% of the maximal aerobic power (MAP). EMG analysis revealed a significant increase in the root mean square (RMS) for all muscles from 70% of the MAP. Gastrocnemius muscles [lateralis gastrocnemius (GasL) and medialis gastrocnemius (GasM)] were the less affected by the increase of PO. Cross-correlation method showed a significant increase in the lag angle values for VM in the last stage compared to the first stage, meaning that the onset of the activation started earlier during the pedaling cycle. Statistical Parametric Mapping (SPM) demonstrated that from 70% MAP, biceps femoris (BF), tibialis anterior (TA), gluteus maximus (GM), and rectus femoris (RF) yielded larger ranges of the crank cycle on which the level of recruitment was significantly increased. This study revealed specific muscular and kinematic coordination for professional cyclists in response to PO increase.