Abstract Previous studies have provided evidence for activation of the human C1 complex by HIV-1, resulting from direct interaction between C1q and the external portion of the viral transmembrane envelope protein, rsgp41. The present study was undertaken to locate more precisely, within C1q and rsgp41, the sites involved in the C1/HIV-1 interaction. Using a solid phase binding assay, we showed that 125I-labeled C1q binding to rsgp41 was dose dependent, saturable, and comparable with binding of C1q to IgG-OVA immune complexes. The globular and, to a lesser extent, the collagen-like regions of C1q both bound to rsgp41. In contrast, the globular region of C1q inhibited the C1q/rsgp41 interaction, whereas the collagen-like region of C1q did not. A series of peptides covering the putative C1q-binding site on gp41 (positions 590-613 of gp160) were synthesized and used as competitors in the C1q-rsgp41-binding assay. Peptide 601-613 (GIWGCSGKLICT) inhibited C1q binding the most efficiently, with 50% inhibition at a concentration of 100 microM. This peptide also inhibited binding of C1q to rsgp36, the protein of HIV-2 homologous to rsgp41. The inhibitory effect of this peptide was dependent in part on the presence of the S-S bridge normally connecting Cys 605 to Cys 611 because reduction of this bond significantly reduced its efficiency. These data suggest that the C1q/HIV-1 interaction involves a site on C1q located within the globular regions, and a major site located within the immunodominant domain of HIV-1, which shares homology with the corresponding region of HIV-2.