Dengue virus (DENV) infects target cells by attaching to various cell receptors, many of which are still unknown. In C6/36 cells (Aedes albopictus cell line), DENV-4 bound to two glycoproteins of 40 and 45 kDa, located on the cell surface. Preincubation of cells with polyclonal antibody against the 45-kDa protein specifically blocked DENV-4 infection of C6/36 cells. The antibody and purified DENV-4 detected the 45-kDa molecule in total extracts from eggs, larvae, and pupae as well as from the midgut, ovary, and salivary glands from adult-stage Aedes aegypti mosquitoes, whereas in malphigian tubules it was absent. This suggests that the distribution of the 45-kDa protein correlates with tissue tropism of DENV infection in mosquitoes. The 45-kDa molecule was not detected in Anopheles albimanus mosquito. The relevance of our findings is discussed from the pathogenetic and vector competence viewpoints.