Dengue virus (DENV) causes a febrile disease, infecting around 50–100 million people annually. The relationship between DENV proteins and host cellular responses during infection is unclear. This study investigated the interaction of host protein disulfide isomerase (PDI) with DENV proteins and role of lipid rafts in viral immunopathogenesis. Host viral protein interactions were studied by co-immunoprecipitation and co-localization. It was found that PDI interacts with DENV nonstructural protein 1 (NS1) intracellularly as well as on the surface in the lipid raft domain. Disruption of this key interaction between PDI and NS1 could be an important therapeutic strategy to block DENV infection.