This study aimed to evaluate how habitat use by medium and large sized mammals was related to different vegetation types in an Atlantic rain-forest reserve in south-eastern Brazil. Our hypotheses were that each vegetation type found in the Vale Natural Reserve (VNR) will present a different mammal species composition, and that mammal species richness and relative abundance will be higher in areas nearest to water. We installed 39 camera traps in the VNR, and used two covariates to characterize the different vegetation types (density of trees and lianas). We also used one spatial covariate: the distance to the nearest water resource. We recorded 26 medium- and large-sized mammal species. The mammalian assemblage was structured by veg- etation type and the proximity of water. The results presented here show that even in a single reserve such as VNR there is a difference in the assemblage composition of medium- and large-sized mammals. Our results also indicate the relevance of water availability when designing protected areas to ensure habitat quality for mammal species. The VNR is one of the major remaining portions of the coastal plain forest of the Atlantic rain forest, and also one of the largest contiguous blocks of this biome.