Chikungunya virus (CHIKV) has recently re‐emerged causing millions of infections in countries around the Indian Ocean. While CHIKV has a broad host cell range and productively infects a number of different cell types, macrophages have been identified as a potential viral reservoir serving to increase the duration of symptoms. To date no CHIKV interacting protein has been characterized and this study sought to identify CHIKV binding proteins expressed on target cell membranes. Two‐dimensional virus overlay identified prohibitin (PHB) as a microglial cell expressed CHIKV binding protein. Co‐localization, co‐immunoprecipitation as well as antibody and siRNA mediated infection inhibition studies all confirmed a role for PHB in mediating internalization of CHIKV into microglial cells. PHB is the first identified CHIKV receptor protein, and this study is evidence that PHB may play a role in the internalization of multiple viruses. J. Med. Virol. 84:1757–1770, 2012. © 2012 Wiley Periodicals, Inc.