NMR mapping of the antigenic determinant recognized by an anti-gp120, human immunodeficiency virus neutralizing antibody.
The 24-amino-acid peptide RP135 (NNTRKSIRIQRGPGRAFVTIGKIG) corresponds in its amino acid sequence to the principal neutralizing determinant of the human immunodeficiency virus type-1, IIIB isolate (HIV-1IIIB, residues 308-331 of the envelope glycoprotein gp120). In order to map the antigenic determinant recognized by 0.5 beta, the complex of RP135 with an anti-gp120 HIV neutralizing antibody, 0.5 beta, which cross reacts with the peptide, was studied by using two-dimensional NMR spectroscopy. A combination of homonuclear Hartmann Hahn two-dimensional experiment and roating-frame Overhauser enhancement spectroscopy of the Fab/peptide complex measured in H2O was used to eliminate the resonances of the Fab and the tightly bound peptide residues and to obtain sequential assignments for those parts of the peptide which retain considerable mobility upon binding. In this manner, a total of 14 residues (Ser6-Thr19) were shown to be part of the antigenic determinant recognized by the antibody 0.5 beta. Lys5 and Ile20 were found to retain considerable mobility in the bound peptide while their amide protons undergo significant change in chemical shift upon binding. This observation suggests that these two residues are at the boundaries of the determinant recognized by the antibody. Competitive binding experiments using truncated peptides strongly support the NMR observations.
Zvi
A
A
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Kustanovich
I
I
Feigelson
D
D
Levy
R
R
Eisenstein
M
M
Matsushita
S
S
Richalet-Sécordel
P
P
Regenmortel
M H
MH
Anglister
J
J
D000595:Amino Acid Sequence; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry*; D015483:HIV Antibodies / Q000737:chemistry*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D018448:Models, Immunological; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D010455:Peptides / Q000138:chemical synthesis; D016415:Sequence Alignment
D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010446:Peptide Fragments; D010455:Peptides; C104122:RP 135 peptide