This study investigates the impact of quadriceps fatigue on lower limb biomechanics during the landing phase of a single-leg vertical jump (SLJ) in 25 amateur male basketball players from Ningbo University. Fatigue was induced through single-leg knee flexion and extension exercises until task failure. Kinematic and dynamic data were collected pre-fatigue (PRF) and post-fatigue (POF) using the Vicon motion capture system and the AMTI force platform and analyzed using an OpenSim musculoskeletal model. Paired sample t-tests revealed significant changes in knee and hip biomechanics under different fatigue conditions, with knee joint angle (p < 0.001), velocity (p = 0.006), moment (p = 0.006), and power (p = 0.036) showing significant alterations. Hip joint angle (p = 0.002), moment (p = 0.033), and power (p < 0.001) also exhibited significant changes. Muscle activation and joint power were significantly higher in the POF condition, while joint stiffness was lower. These findings suggest that quadriceps fatigue leads to biomechanical adjustments in the knee and hip joints, which may increase the risk of injury despite aiding in landing stability.