Coarse-scale studies that focus on species distributions and richness neglect heterogeneity that may be present at finer scales. Studies of arthropod assemblage structure at fine (1 x 1 km) scales are rare, but important, because these are the spatial levels at which real world applications are viable. Here we investigate fine-scale variation in spider assemblages, comparing five representative vegetation types in the western Soutpansberg, Limpopo Province, South Africa. We assess these vegetation types in terms of their family and species composition, as well as levels of endemicity, relating these differences with vegetation structure. We inventoried 297 species (49 families) in an area less than 450 ha, as part of South African National Survey of Arachnida. Analysis of the results suggests that endemic taxa are associated with Tall Forest and, to a lesser extent, Woodland. The Woodland had the highest species diversity, and much of the variation observed in spider assemblage structure is explained by these two vegetation types. Based on vegetation structure variables that explained significant variation in spider assemblages, human influence through bush encroachment will result in a change of spider assemblages to that of Short Forest and Mosaic Woodland vegetation types, with implications for biodiversity maintenance and heterogeneity.