Characterization of interaction between C-domain on HIV-1 gp41 and the putative receptor protein p62

Based on the fact that the binding of HIV-1 gp41 to the putative cellular receptor protein p62 could be inhibited by the C-domain peptide of gp41, we wanted to confirm the interaction of the C-domain with p62 and to characterize the receptor-binding site on the C-domain. We attempted to isolate the putative receptor protein p62 from cell lysates of the human B cell line Raji by affinity chromatography using sepharose-columns which were conjugated with three different peptides of the C-domain respectively. A protein of 62 kDa was isolated by peptide (NP2)-sepharose-column, while none of the proteins was identified in eluates of the other two overlapped peptides of C-domain, indicating that HIV-1 gp41 by the region aa635-664 of C-domain binds to the putative receptor protein p62. Besides, CD spectroscopy analysis revealed that only a NP2 peptide could induce significant conformational change of P62. In addition, the interaction between P62 and three peptides of the C-domain was characterized by the surface plasma resonance (SPR) measurement. It was indicated that only the NP2 peptide significantly inhibited the interaction between rsgp41 and the putative receptor P62, confirming that the protein p62 may serve as a potential receptor for gp41 binding, and the peptide NP2 contains an integrate binding site for gp41 binding to p62.