The purpose of this study was to compare the relationship between surface electromyography (EMG) and knee joint angle of the vastus intermedius muscle (VI) with the synergistic muscles in the quadriceps femoris (QF) muscle group. Fourteen healthy men performed maximal voluntary contractions during isometric knee extension at four knee joint angles from 90°, 115°, 140°, and 165° (180° being full extension). During the contractions, surface EMG was recorded at four muscle components of the QF muscle group: the VI, vastus lateralis (VL), vastus medialis (VM), and rectus femoris (RF) muscles. The root mean square of the surface EMG at each knee joint angle was calculated and normalized by that at a knee joint angle of 90° for individual muscles. The normalized RMS of the VI muscle was significantly lower than those of the VL and RF muscles at the knee joint angles of 115° and 165° and those of the VL, VM, and RF muscles at the knee joint angle of 140° (P<0.05). The present results suggest that the neuromuscular activation of the VI muscle is regulated in a manner different from the alteration of the knee joint angle compared with other muscle components of the QF muscle group.