Molecular recognition in the HIV-1 capsid/cyclophilin a complex

The HIV-1 capsid protein (CA) makes an essential interaction with the human peptidyl prolyl isomerase, cyclophilin A (CypA), that results in packaging of CypA into the virion at a CA to CypA stoichiometry of similar to 10:1. The 231 amino acid residue capsid protein is composed of an amino-terminal CypA binding domain (1 to similar to 151; CA(151)) and a carboxylterminal dimerization domain (similar to 151 to 231). We find that CypA binds dimeric CA and monomeric CA(151) with identical intrinsic affinities (K-d = 16(+/-4)mu M) This result demonstrates that capsid dimerization and cyclophilin A binding are not thermodynamically coupled and suggests that the substoichiometric ratio of CypA in the HIV-1 virion results from the intrinsic stability of the CA/CypA complex. Ln the known co-crystal structure of the CA(151)/CypA complex, CypA binding is mediated exclu sirely by an exposed capsid loop that spans residues Pro85 to Pro93. The energetic contributions to CypA binding were quantified for each residue in this loop, and the results demonstrate that the Gly89-Pro90 dipeptide is the primary cycloyhilin A recognition motif, with Pro85, Val86, His87, Ala88, and Pro93 also making energetically favorable contacts. These studies reveal that the active site of CypA, which can catalyze the isomerization of proline residues in vitro, also functions as a sequence-specific, protein-binding motif in HIV-1 replication. (C) 1997 Academic Press Limited.