The aim of this study is to investigate the differences in muscle contractile properties measured by the TMG method between top-level football players of different playing positions. The sample consisted of 57 football players from the Serbian Super League, divided into three groups: defenders-DF, midfielders-MF, and forwards-FW. Muscles included in the study were the Rectus Femoris (RF), Vastus Medialis (VM), Vastus Lateralis (VL), Biceps Femoris (BF), and Semitendinosus (ST) of the right (R) and left (L) leg. The TMG parameters used in this study were contraction time (Tc), delay time (Td), relaxation time (Tr), maximal displacement (Dm), and sustain time (Ts). The ANOVA results showed that differences in TMG parameters between top-level football players of different playing positions are small and exist only in the Tr of RF (F = 4.658, = 0.014), BF (F = 4.433, = 0.016), and ST muscle (F = 3.808, = 0.028), and the Tc (F = 3.214, = 0.048) and Td (F = 3.705, = 0.031) of the VM muscle. All differences were detected between DF and FW players, and all differences were in the left (non-dominant) leg. The results obtained in this study indicate that the training and selection process in football, from the aspect of functional and mechanical muscle properties, should be similar for all players, regardless of playing position. It has been shown that TMG has relatively low sensitive strength for detecting differences between football players of different playing positions.