title: Crystal structure of the neutralizing Llama V(HH) D7 and its mode of HIV-1 gp120 interaction.
abstract: HIV-1 entry into host cells is mediated by the sequential binding of the envelope glycoprotein gp120 to CD4 and a chemokine receptor. Antibodies binding to epitopes overlapping the CD4-binding site on gp120 are potent inhibitors of HIV entry, such as the llama heavy chain antibody fragment V(HH) D7, which has cross-clade neutralizing properties and competes with CD4 and mAb b12 for high affinity binding to gp120. We report the crystal structure of the D7 V(HH) at 1.5 A resolution, which reveals the molecular details of the complementarity determining regions (CDR) and substantial flexibility of CDR3 that could facilitate an induced fit interaction with gp120. Structural comparison of CDRs from other CD4 binding site antibodies suggests diverse modes of interaction. Mutational analysis identified CDR3 as a key component of gp120 interaction as determined by surface plasmon resonance. A decrease in affinity is directly coupled to the neutralization efficiency since mutations that decrease gp120 interaction increase the IC50 required for HIV-1 IIIB neutralization. Thus the structural study identifies the long CDR3 of D7 as the key determinant of interaction and HIV-1 neutralization. Furthermore, our data confirm that the structural plasticity of gp120 can accommodate multiple modes of antibody binding within the CD4 binding site.
authors.affiliation: Unit of Virus Host Cell Interactions (UVHCI), UMI 3265, Université Joseph Fourier-EMBL-CNRS, Grenoble, France.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hinz
authors.forename: David
authors.initials: D
authors.lastname: Lutje Hulsik
authors.forename: Anna
authors.initials: A
authors.lastname: Forsman
authors.forename: Willie Wee-Lee
authors.initials: WW
authors.lastname: Koh
authors.forename: Hassan
authors.initials: H
authors.lastname: Belrhali
authors.forename: Andrea
authors.initials: A
authors.lastname: Gorlani
authors.forename: Hans
authors.initials: H
authors.lastname: de Haard
authors.forename: Robin A
authors.initials: RA
authors.lastname: Weiss
authors.forename: Theo
authors.initials: T
authors.lastname: Verrips
authors.forename: Winfried
authors.initials: W
authors.lastname: Weissenhorn
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015704:CD4 Antigens / Q000276:immunology; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000276:immunology; D004252:DNA Mutational Analysis; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000187:drug effects; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000276:immunology* / Q000494:pharmacology; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry* / Q000276:immunology* / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009500:Neutralization Tests; D011485:Protein Binding / Q000187:drug effects; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D012997:Solvents; D040681:Structural Homology, Protein; D013499:Surface Properties / Q000187:drug effects
chemical_list: D015704:CD4 Antigens; D022801:Complementarity Determining Regions; D015699:HIV Envelope Protein gp120; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D050505:Mutant Proteins; D012997:Solvents
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