The major core protein (p25) of the human immunodeficiency virus type 1 (HIV-1) was characterized by two-dimensional-gel isoelectric focusing. The p25 detectable in HIV-1-infected cells is composed of four species with related isoelectric points. This is due in part to the phosphorylated state of p25. The four species of p25 are expressed on the cell surfaces of infected cells, but only the two most basic species are incorporated into the HIV-1 virion. These findings emphasize the importance of p25 in understanding infection with HIV and might have implications for the development of vaccines.