The southeastern United States (SE-US) has undergone one of the highest rates of landscape changes in the country due to changing demographics and land use practices over the last few decades. Increasing evidence indicates that these changes have impacted mesoscale weather patterns, biodiversity and water resources. Since the Southeast has one of the highest rates of land productivity in the nation, it is important to monitor the effects of such changes regularly. Here, we propose a remote sensing based methodology to estimate regional impacts of urban land development on ecosystem structure and function. As an indicator of ecosystem functioning, we chose net primary productivity (NPP), which is now routinely estimated from the MODerate resolution Imaging Spectroradiometer (MODIS) data. We used the MODIS data, a 1992 Landsat-based land cover map and nighttime data derived from the Defense Meteorological Satellite Program's Operational Linescan System (DMSP/OLS) for the years 1992/1993 and 2000 to estimate the extent of urban development and its impact on NPP. The analysis based on the nighttime data indicated that in 1992/1993, urban areas amounted to 4.5% of the total land surface of the region. In the year 2000, the nighttime data showed an increase in urban development for the southeastern United States of 1.9%. Estimates derived from the MODIS data indicated that land cover changes due to urban development that took place during the 1992–2000 period reduced annual NPP of the southeastern United States by 0.4%. Despite the uncertainties in sensor fusion and the coarse resolution of the data used in this study, results show that the combination of MODIS products such as NPP with nighttime data could provide rapid assessment of urban land cover changes and their impacts on regional ecosystem resources.