Binding thermodynamics of the N-terminal peptide of the CCR5 coreceptor to HIV-1 envelope glycoprotein gp120.

The initial events of HIV-1 cell infection involve the sequential binding of the viral envelope glycoprotein gp120 to the cellular CD4 receptor and the coreceptor, usually CCR5 or CXCR4. Binding to the coreceptor triggers the chain of events that culminates with the entry of the virus into the cell. In this process, the interaction of gp120 with the tyrosine-sulfated N-terminus of CCR5 is critical; however, this interaction has never been characterized at a quantitative or thermodynamic level. Here, we present the first thermodynamic analysis of the interaction of gp120 with the N-terminal peptide of the CCR5 coreceptor. Microcalorimetric titrations demonstrate that measurable binding of S22 peptide, a 22-amino acid tyrosine-sulfated peptide corresponding to the CCR5 N-terminus, requires prior binding of CD4 to gp120. The S22 peptide binds to the gp120-CD4 complex with a binding affinity of 4.5 x 10(5) M(-1) (K(d) = 2.2 microM) in an enthalpically and entropically favorable process. An identical peptide lacking the sulfated tyrosine residues is unable to bind the gp120-CD4 complex. These results indicate that the sulfated tyrosines contribute close to -3.5 kcal/mol to the Gibbs energy of binding. Furthermore, the S22 peptide is a competitive inhibitor of the 17b HIV-1 neutralizing antibody, which is known to bind to the CCR5 coreceptor site in gp120. Together, these results point toward compounds containing sulfated aromatic groups as potential inhibitors of viral entry. In analogy to existing inhibitors that bind to the CCR5 coreceptor directly, these compounds will accomplish the same result by binding to the coreceptor site in gp120.