As urban habitats vary in composition and structure along the urban to rural gradient, different degrees of urbanization likely result in a diversity of landscape responses from wildlife. We investigated this relationship with the Virginia opossum (Didelphis virginiana), an urban adapted species that is both common and understudied in highly metropolitan landscapes. We investigated which landscape factors affect opossum occupancy, colonization, extinction, and detection by using a large system of motion-triggered camera traps in the Chicago metropolitan area over 10 seasons from spring 2010 to summer 2012. Opossum patch occupancy rates were highest near natural water sources regardless of urbanization, whereas occupancy rates in patches ≥1000 m from natural water sources decreased with increasing urbanization. Our results suggest opossums have relaxed habitat needs at intermediate levels of disturbance, as the ability to locate anthropogenic water sources may allow them to occupy previously uninhabitable patches.