The team tactical system and distribution of the football players on the pitch is considered fundamental in team performance. The present study used time-motion analysis and triaxial-accelerometers to obtain new insights about the impact of different tactical systems (1-4-5-1 and 1-3-5-2) on physical performance, across different playing positions, in a professional football team. Player performance data in fifteen official home matches was collected for analysis. The sample included twenty-two players from five playing positions (centre backs: n = 4; full-back/wide midfielder/ wing-back: n = 9; centre midfielder: n = 6 and centre forward: n = 3), making a total of 108 match observations. A novel finding was that general match physical demands do not differ considerably between these tactical formations, probably because match-to-match variability (variation of players' running profile from match-to-match) might be higher than the differences in physical performance between tactical systems. However, change of formation had a different impact across playing positions, with centre backs playing in 1-4-5-1 performing significant more HIRcounts than in 1-3-5-2 (p = 0.031). Furthermore, a medium effect size (r = 0.33) was observed in HIRdist, with wide players covering higher distances when playing in 1-3-5-2 than in 1-4-5-1. These findings may help coaches to develop individualised training programs to meet the demands of each playing position according to the tactical system adopted.