Habitat loss and fragmentation are widely recognized as among the most important threats to global biodiversity. New analytical approaches are providing improved ability to predict the effects of landscape change on population connectivity at vast spatial extents. This paper presents an analysis of population connectivity for three species of conservation concern [swift fox (Vulpes velox); lesser prairie-chicken (Tympanuchus pallidicinctus); massasuaga (Sistrurus catenatus)] across the American Great Plains region. We used factorial least-cost path and resistant kernel analyses to predict effects of landscape conditions on corridor network connectivity. Our predictions of population connectivity provide testable hypotheses about the location of core habitats, corridors, and barriers to movement. The results indicate that connectivity is more sensitive to a species’ dispersal ability than variation in landscape resistance to movement. Thus, it may prove difficult to optimize conservation strategies to maintain population connectivity for multiple species with disparate dispersal abilities and independent distributions.