This study was designed to evaluate the way in which changes in vertical jaw opening affected the relative contributions of various masticatory muscles to bite force production. EMG activity was recorded simultaneously from the masseter, and anterior, middle, and posterior temporalis muscles, during controlled isometric biting at different force levels and vertical jaw openings. EMG-force characteristics were compared between muscles and bite openings. All but the posterior temporalis muscle displayed significant increases in muscle activity with increased bite force production; the masseter muscle demonstrated the largest activity increments. Statistically significant changes in muscle function due to jaw opening were demonstrated only for the masseter muscle, though similar trends were observed for the anterior and middle temporalis muscles. Minimum increases in muscle activity associated with increases in bite force occurred between 9 and 11 mm of opening, measured at the first molar, for all three muscle groups. The results of this study suggest that changes in masticatory muscle length resulting from vertical jaw opening cause alterations in muscle contractile properties, but the relative contributions of various masticatory muscles toward bite force production may also be affected by biomechanical factors and neural control adaptations.