Accessibility is the ease of reaching opportunities (goods, services, activities, and destinations). Accessibility of desirable locations such as households and commercial locations, is typically scaffolded by land use patterns and transportation infrastructure. It can reflect people’s travel convenience, cities’ viability, sustainability, and mitigate the negative effects on the environment and public safety. Consequently, it is recognized as a fundamental principle in urban sustainable development policies worldwide. In the literature, most of the studies have used a static or partially dynamic approach with a single mode such as a car or public transportation by using conventional models. These “static” models assume that household locations are static and that transportation supply and opportunities for social practice activities are fixed in time and space, which can lead to biased or even misleading assumptions in accessibility models. Therefore, the aim of this study is to evaluate the impact of dynamic spatial accessibility through Mode-Dependent Accessibility (MDA) on the location choice behaviors of urban activities such as households and commercialin the City of Wuhan, China. This study employed the Mode-Dependent Travel Demand Model (M-TDM) to measure the impact of short-term MDA on household and commercial activities for the years 2012 and 2015. Additionally, an integrated spatial economic (ISE) model such as PECAS (Production, Exchange, Consumption, Allocation, System) in order to investigate location preferences of urban activities over space and time. Regarding household and commercial location choice, the ISE modeling results revealed that households and commercial activities are sensitive to MDA, especially using transit. The ISE method predicted that the R2 for household and commercial location choice models was 0.84 to 0.90 for transit-based accessibility, whereas the R2 for logsum-based static models was 0.48 to 0.72. In addition, their findings suggest that highly accessible locations that are well served by auto are more appealing for household and commercial activities. The findings of this study will help urban planners, transportation planners, and policymakers take into account the dynamic nature of short-term MDA when zoning and allocating urban activities and public amenities, instead of using static accessibility.