title: Isolate-specific differences in the conformational dynamics and antigenicity of HIV-1 gp120.
abstract: The HIV-1 envelope glycoprotein (Env) mediates viral entry into host cells and is the sole target of neutralizing antibodies. Much of the sequence diversity in the HIV-1 genome is concentrated within Env, particularly within its gp120 surface subunit. While dramatic functional diversity exists among HIV-1 Env isolates-observable even in the context of monomeric gp120 proteins as differences in antigenicity and immunogenicity-we have little understanding of the structural features that distinguish Env isolates and lead to isolate-specific functional differences, as crystal structures of truncated gp120 "core" proteins from diverse isolates reveal a high level of structural conservation. Because gp120 proteins are used as prospective vaccine immunogens, it is critical to understand the structural factors that influence their reactivity with antibodies. Here, we studied four full-length, glycosylated gp120 monomers from diverse HIV-1 isolates by using small-angle X-ray scattering (SAXS) to probe the overall subunit morphology and hydrogen/deuterium-exchange with mass spectrometry (HDX-MS) to characterize the local structural order of each gp120. We observed that while the overall subunit architecture was similar among isolates by SAXS, dramatic isolate-specific differences in the conformational stability of gp120 were evident by HDX-MS. These differences persisted even with the CD4 receptor bound. Furthermore, surface plasmon resonance (SPR) and enzyme-linked immunosorbance assays (ELISAs) showed that disorder was associated with poorer recognition by antibodies targeting conserved conformational epitopes. These data provide additional insight into the structural determinants of gp120 antigenicity and suggest that conformational dynamics should be considered in the selection and design of optimized Env immunogens.
authors.affiliation: Department of Global Health.
authors.forename: Thaddeus M
authors.initials: TM
authors.lastname: Davenport
authors.forename: Miklos
authors.initials: M
authors.lastname: Guttman
authors.forename: Wenjin
authors.initials: W
authors.lastname: Guo
authors.forename: Brad
authors.initials: B
authors.lastname: Cleveland
authors.forename: Maria
authors.initials: M
authors.lastname: Kahn
authors.forename: Shiu-Lok
authors.initials: SL
authors.lastname: Hu
authors.forename: Kelly K
authors.initials: KK
authors.lastname: Lee
mesh_terms: D016915:AIDS Vaccines; D057134:Antibodies, Neutralizing; D000956:Antigens, Viral / Q000276:immunology* / Q000378:metabolism; D015704:CD4 Antigens / Q000378:metabolism; D041961:Deuterium Exchange Measurement; D015195:Drug Design; D000939:Epitopes / Q000276:immunology*; D006031:Glycosylation; D015483:HIV Antibodies / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D011487:Protein Conformation; D053838:Scattering, Small Angle; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020349:Surface Plasmon Resonance
chemical_list: D016915:AIDS Vaccines; D057134:Antibodies, Neutralizing; D000956:Antigens, Viral; D015704:CD4 Antigens; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120
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