Both the medial collateral ligament (MCL) and the anterior cruciate ligament (ACL) are reported to prevent valgus instability of the knee. In this study, the anatomical mechanisms by which these ligaments prevent valgus instability were experimentally investigated. The valgus rotation angle and the magnitude of the medial joint space opening were measured in six cadaveric knees, using biplanar photography before and after the MCL and/or the ACL were severed. A significant increase in the valgus rotation angle and a large medial joint space opening were observed when the MCL was severed. An increase in the valgus rotation angle was also observed when the ACL was severed, but only a small medial joint space opening was present. The increase in the valgus rotation angle after ACL severance was nearly parallel to the increase in the internal rotation of the tibia. Thus, we concluded that both ligaments function to prevent valgus instability, but that the anatomical reasons for their function are different. The MCL prevents valgus instability by stopping an opening in the medial joint space. The ACL, on the other hand, prevents the internal rotation of the tibia. When the ACL is severed, the internal rotation increases, and causes the valgus rotation angle to also increase, despite the presence of only a small medial joint space opening.