The maintenance of viable carnivore populations along human-dominated landscapes depends on the understanding of the species requirements that are critical to the design of global actions focused on their conservation. Using resource selection function and a spatial analysis approach, we evaluated large carnivore habitat use in a disturbed landscape by studying pumas in a rapidly developing region in Southeast Brazil. Pumas had a mean home-range size of 203.7 ± 39.8 km2 and showed pronounced territorial behavior through intrasexual overlap (males 22% and females 60%). Our results also revealed a strong habitat association with forest vegetation both at population and individual levels, followed by pasture with shrubs, a type of anthropized habitat that offers cover and prey; the use of this type of landcover was more frequent during night hours. Our study is the first robust home-range estimation of pumas in Brazil and fills a gap in puma’s ecology knowledge in anthropized tropical environments, highlighting the importance of natural habitat patches to this predator along human-modified areas and reinforcing the species demand for vast and forested areas to thrive. In areas where conservation goals include the maintenance or increase in large predator populations, identifying the limits imposed by anthropogenic landscape changes, and ensuring that these are not exceeded, is paramount for conservation initiatives.