The Paddlefish Polyodon spathula is an ancient species that is native to the Mississippi River and its larger tributaries. This species exhibits a unique combination of morphology and life history characteristics that leaves it vulnerable to river modifications and potential overexploitation, which has led to population declines in portions of the historic range. Concern regarding unknown exploitation rates from recreational and commercial fisheries has increased in recent decades, and the Convention on International Trade of Endangered Species is now requesting information from state agencies regarding the sustainability of commercially harvested Paddlefish populations. The Missouri Department of Conservation addressed this, in part, through the implementation of a 5-year mark-recapture study to estimate exploitation and nonreporting rates, evaluate movement patterns, and provide a baseline estimate of abundance for Paddlefish in the Mississippi River basin. A total of 2,224 Paddlefish were jaw-banded, and 102 were also implanted with transmitters in the Mississippi River basin within the borders of Missouri. Of those, 139 Paddlefish were caught by commercial or recreational anglers and 92 were harvested, resulting in an average annual exploitation estimate of 2.5% (SE = 0.6). Jaw band returns and telemetry information demonstrated that Paddlefish moved great distances (maximum 2,359 km) and crossed many regulatory boundaries (14 states). Despite relatively low exploitation rate estimates, a precautionary adjustment of regulations is advised to protect Paddlefish through maturation and ensure sustainability. In addition, regulations for this species should be assessed across the entire Mississippi River basin, as they differ within and between regulatory authorities. A combination of population monitoring (e.g., exploitation and population dynamics) and telemetry efforts have potential to help inform future basin-wide management approaches.