We have identified a cellular protein from a continuous mosquito cell line (C6/36) that appears to play a significant role in the attachment of Venezuelan equine encephalitis (VEE) virus to these cells. VEE virus bound to a 32-kDa polypeptide present in the C6/36 plasma membrane fraction, and binding to this polypeptide was dose dependent and saturable and competed with homologous and heterologous alphaviruses. These observations suggest that this polypeptide binds virus via a receptor-ligand interaction. The 32-kDa polypeptide was expressed on the surfaces of C6/36 cells, and monoclonal antibodies directed against either this cell polypeptide or the VEE virus E2 glycoprotein, which is thought to be the viral attachment protein, interfered with virus attachment. Collectively, these data provide evidence suggesting that the 32-kDa polypeptide serves as a receptor for VEE virus infection of cells. We have characterized this cell polypeptide as a laminin-binding protein on the basis of its ability to interact directly with laminin as well as its immunologic cross-reactivity with the high-affinity human laminin receptor.The ability of viruses to bind to host cells is controlled by the interaction of viral attachment proteins with cellular receptor proteins (CRP). The interaction between viral attachment proteins and CRP is thought to influence tissue tropism and/or host range for many viruses (12,14,20,22,36,39,40). This phenomenon is particularly evident for some mammalian viruses, for which the ability of the virus to bind to distinct host cells (1,4,9,30) plays an important role in viral pathogenesis (5).The Venezuelan equine encephalitis (VEE) virus complex is a group of serologically related alphaviruses in the family Togaviridae. In nature, these viruses are transmitted between susceptible vertebrate hosts by a variety of mosquito species (40). Virions are spherical (60 to 65 nm in diameter) and are composed of an icosahedral nucleocapsid containing a positivestranded RNA genome. The nucleocapsid is surrounded by a lipid envelope containing two structural glycoproteins, E1 and E2. Certain domains of these two glycoproteins are exposed on the surface of virions and are responsible for the interaction of virus with neutralizing antibody. It has been reported previously that at least one of the E2 domains modulates the attachment of virus to cells (35).VEE virus is typical of arboviruses in that transmission, dissemination, and amplification require cyclic passage through vertebrate and invertebrate hosts. The fact that infections of unrelated organ systems occur in such dissimilar hosts suggests either that these viruses utilize different strategies for attachment to different host cells or that they attach to cellular components which are highly conserved among phylogenetically distinct species. An understanding of the molecular basis of host specificity, vector competence, and tissue tropisms of VEE virus would be greatly facilitated by a better understanding of virus-receptor interactions.Studies have shown th...