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 approximately 10:1. The 231 amino acid residue capsid protein is composed of an amino-terminal CypA binding domain (1 to approximately 151; CA151) and a carboxyl-terminal dimerization domain (approximately 151 to 231). We find that CypA binds dimeric CA and monomeric CA151 with identical intrinsic affinities (K[d] = 16(+/-4) microM). 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. In the known co-crystal structure of the CA151/CypA complex, CypA binding is mediated exclusively 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 cyclophilin 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.
Yoo
S
S
Department of Biochemistry, University of Utah, Salt Lake City 84132, USA.
Myszka
D G
DG
Yeh
C
C
McMurray
M
M
Hill
C P
CP
Sundquist
W I
WI
D000591:Amino Acid Isomerases / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D002151:Calorimetry; D002213:Capsid / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D019281:Dimerization; D015683:Gene Products, gag / Q000378:metabolism*; D015497:HIV-1 / Q000145:classification / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D019696:Peptidylprolyl Isomerase; D011485:Protein Binding; D011994:Recombinant Proteins; D012542:Scattering, Radiation; D012996:Solutions; D013057:Spectrum Analysis; D013816:Thermodynamics; D017186:Titrimetry; D014779:Virus Replication
D002352:Carrier Proteins; D015683:Gene Products, gag; D011994:Recombinant Proteins; D012996:Solutions; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase