Previous research has suggested that personality traits of the Five Factor Model play a role in worker's response to workload. The aim of this study was to investigate the association of personality traits of first responders with their perceived workload in real-life tasks. A flying column of 269 police officers completed a measure of subjective workload (NASA-Task Load Index) after intervention tasks in a major public event. Officers' scores on a measure of Five Factor Model personality traits were obtained from archival data. Linear Mixed Modeling was used to test the direct and interaction effects of personality traits on workload scores once controlling for background variables, task type and workload source (mental, temporal and physical demand of the task, perceived effort, dissatisfaction for the performance and frustration due to the task). All personality traits except extraversion significantly interacted at least with one workload source. Perceived workload in flying column police officers appears to be the result of their personality characteristics interacting with the workload source. The implications of these results for the development of support measures aimed at reducing the impact of workload in this category of workers are discussed.