Understanding the in-game demands placed on athletes may allow practitioners to design improved training protocols to prepare athletes for competitive demands. This study aimed to quantify the competitive movement demands of professional American football athletes and to determine any inter-positional differences that may exist. Player tracking data were collected from 2018 to 2020 regular season games of the National Football League. Distance, maximum velocity, high-velocity efforts and distance, and acceleration and deceleration efforts and distance were used to evaluate competitive movement demands. To determine position-specific demands, each player was classified by their designated position, and velocity data from competitive games were used to develop position-specific velocity thresholds. One-way ANOVA and post hoc Bonferroni statistical analysis were used to determine inter-positional difference. Significant (p < 0.05) positional differences were found for all load metrics with respect to competitive game demands. Generally, wide receivers and defensive backs had faster maximum velocities, higher distances, and more acceleration and deceleration efforts and distance than other positions. Linebackers accumulated the most high-velocity efforts and distance. Lineman had the lowest values for all assessed metrics. These findings may assist the performance staff in developing improved training and return-to-play protocols with the aim of improving player performance and mitigating injury.