In recent years, accessibility has been considered a vital policy objective in the development of a sustainable transportation system. Accessibility can make a significant contribution to quality of life. The accessibility of desirable locations, such as households and commercial locations, is usually underpinned by land use patterns and transportation infrastructure. The accessibility of different activities is largely determined by the spatial distribution of activities and associated transport networks in a metropolitan area. In order to examine how location choice behaviors of urban activities influence urban forms and land use patterns, accessibility has been used extensively to consider the impact of both the spatial distribution of opportunities (e.g., employment or services) and the transport cost of reaching them. However, in most cases, only those static/aggregate accessibility terms that are represented by the “logsum” of all available transport modes have traditionally been used in urban transport planning and modeling analysis. In this study, according to urban activities, several Integrated Spatial Economic (ISE) models i.e., PECAS (Production, Exchange, Consumption, Allocation, System) models were developed to study the relationship between dynamic, Time-Dependent Accessibility (TDA) and the location choice behavior of urban activities in the City of Wuhan, China. The developed models were then used as tools to investigate the impact of dynamic/disaggregate short-term TDA on location choice behaviors of various urban activities such as households and commercial. Regarding the household location choice, the ISE modeling results revealed that urban households living in the downtown area of the City of Wuhan were sensitive to TDA to employment centers, especially during the morning peak time. In addition, commercial services prefer locations that offer a high level of accessibility during off-peak times. Based on the results of this study, it is recommended that planning exercises, such as the development of zoning and the allocation of urban activities and public facilities, pay more attention to dynamic, short-term TDA, which is essential for urban sectors to carry out daily activities, than their static, composite accessibility counterparts.