Dissociation of the CD4 and CXCR4 Binding Properties of Human Immunodeficiency Virus Type 1 gp120 by Deletion of the First Putative Alpha-Helical Conserved Structure

Abstract

To evaluate conserved structures of the surface gp120 subunit (SU) of the human immunodeficiency virus type 1 (HIV-1) envelope in gp120-cell interactions, we designed and produced an HIV-1 IIIB (HXB2R) gp120 carrying a deletion of amino acids E61 to S85. This sequence corresponds to a highly conserved predicted amphipathic alpha-helical structure located in the gp120 C1 region. The resultant soluble mutant with a deleted alpha helix 1 (gp120 ΔαHX1) exhibited a strong interaction with CXCR4, although CD4 binding was undetectable. The former interaction was specific since it inhibited the binding of the anti-CXCR4 monoclonal antibody (12G5), as well as SDF1α, the natural ligand of CXCR4. Additionally, the mutant gp120 was able to bind to CXCR4 + /CD4 − cells but not to CXCR4 − /CD4 − cells. Although efficiently expressed on cell surface, HIV envelope harboring the deleted gp120 ΔαHX1 associated with wild-type transmembrane gp41 was unable to induce cell-to-cell fusion with HeLa CD4 + cells. Nevertheless, the soluble gp120 ΔαHX1 efficiently inhibited a single round of HIV-1 LAI infection in HeLa P4 cells, with a 50% inhibitory concentration of 100 nM. Our data demonstrate that interaction with the CXCR4 coreceptor was maintained in a SUgp120 HIV envelope lacking αHX1. Moreover, in the absence of CD4 binding, the interaction of gp120 ΔαHX1 with CXCR4 was sufficient to inhibit HIV-1 infection.