A synthetic conformational epitope from the C4 domain of HIV Gp120 that binds CD4.
The fourth conserved domain of the human immunodeficiency virus type 1 (HIV-1) envelope, the C4 region of glycoprotein 120 (gp120), is believed to be a major part of gp120 that is necessary for binding to CD4. Recently, we found that C4 in gp120 is probably an alpha-helix, because antibodies made against helical constructs of C4 react with native and recombinant gp120 but antibodies against linear C4 constructs do not. For the present study, we performed experiments to determine, first, if CD4 could bind to the helical C4 constructs and, second, if the binding was comparable with CD4 binding to gp120. Immobilized helical constructs derived from the C4s from HIV-1 and HIV-2 bound biotinylated recombinant CD4 with Kd values of 8.59 nM and 14.59 nM, respectively. Recombinant soluble CD4 inhibited the binding of biotinylated CD4 to the C4 construct from HIV-1 with a Kd of 9.88 nM, and recombinant gp120 blocked the binding of CD4 to the immobilized helical construct from C4 of HIV-1 with a Kd of 8.08 nM. The C4 peptide-(419-436) from HIV-1 (KIKQIINMWQEVGKAMYA-NH2) blocked CD4 binding to gp120 but only in a buffer containing 0.03% Brij 35 where the peptide displayed 17 +/- 1% alpha-helix; without the Brij 35, peptide-(419-436) displayed no helical content. The Kd for the peptide-(419-436) blocking CD4 binding to gp120 in Brij 35-containing buffer was found to be 42 microM. These results indicate that C4 constructs from HIV-1 and HIV-2 do bind CD4, but the constructs must display an alpha-helical conformation to do so. In addition, the results reported here will provide answers to key questions about structural requirements for HIV vaccines and therapeutics that hinge on understanding the molecular nature of the gp120-CD4 interaction as the first step in the HIV infection process.
Robey
F A
FA
Peptide and Immunochemistry Unit, NIDR, National Institutes of Health Bethesda, Maryland 20892, USA.
Harris-Kelson
T
T
Robert-Guroff
M
M
Batinić
D
D
Ivanov
B
B
Lewis
M S
MS
Roller
P P
PP
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D015704:CD4 Antigens / Q000378:metabolism*; D002942:Circular Dichroism; D000939:Epitopes / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000378:metabolism*; D015497:HIV-1 / Q000276:immunology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
D015704:CD4 Antigens; D000939:Epitopes; D015699:HIV Envelope Protein gp120