Partial Enzymatic Deglycosylation Preserves the Structure of Cleaved Recombinant HIV-1 Envelope Glycoprotein Trimers

Depetris, Rafael S.; Julien, Jean-Philippe; Khayat, Reza; Lee, Jeong Hyun; Pejchal, Robert; Katpally, Umesh; Cocco, Nicolette; Kachare, Milind D; Massi, Evan; David, Kathryn B.; Cupo, Albert; Marozsan, Andre J.; Olson, William C.; Ward, Andrew B.; Wilson, Ian A.; Sanders, Rogier W.; Moore, John P.

doi:10.1074/jbc.m112.371898

Cited by 50 publications

(61 citation statements)

References 83 publications

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“…Accordingly, such a quaternary epitope might elicit a particularly productive response from the host's humoral immune system. Here, we show that the BG505 SOSIP.664 gp140 trimer not only binds PG9, but is also more thermally stable than gp120 monomers (50, 51) and previously described SOSIP gp140 trimers (34). Studies comparing how neutralizing and nonneutralizing Abs recognize the CD4bs on Env highlight the importance of epitope presentation in the context of the functional trimer (50).…”

Section: Discussionmentioning

confidence: 89%

“…Thus, the introduced glycan moiety has quantitatively similar adverse effects on both trimer binding and virus neutralization. Together, these results indicate that PG9 LCDR2, the site of the introduced The EM reconstruction of the unliganded BG505 SOSIP.664 gp140 trimer showed a slightly more compact structure than the corresponding unliganded trimer based on the KNH1144 sequence (34) (Fig. 4A and Fig.…”

Section: Molecular Structure Revealed By Negative Stain Single Particmentioning

confidence: 94%

“…These trimers are based on the BG505 Clade A sequence, cleaved at the gp120/gp41 junction, stabilized by SOSIP mutations, and truncated at residue 664 of the gp41 ectodomain (34)(35)(36)(37). Several biochemical and biophysical techniques, alone and in combination, clearly show that only a single PG9 fragment antigen-binding (Fab) binds each BG505 SOSIP.664 glycoprotein 140 (gp140) trimer.…”

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confidence: 99%

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Asymmetric recognition of the HIV-1 trimer by broadly neutralizing antibody PG9

Julien

Lee

Cupo

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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PG9 is the founder member of an expanding family of glycandependent human antibodies that preferentially bind the HIV (HIV-1) envelope (Env) glycoprotein (gp) trimer and broadly neutralize the virus. Here, we show that a soluble SOSIP.664 gp140 trimer constructed from the Clade A BG505 sequence binds PG9 with high affinity (∼11 nM), enabling structural and biophysical characterizations of the PG9:Env trimer complex. The BG505 SOSIP.664 gp140 trimer is remarkably stable as assessed by electron microscopy (EM) and differential scanning calorimetry. EM, small angle X-ray scattering, size exclusion chromatography with inline multiangle light scattering and isothermal titration calorimetry all indicate that only a single PG9 fragment antigen-binding (Fab) binds to the Env trimer. An ∼18 Å EM reconstruction demonstrates that PG9 recognizes the trimer asymmetrically at its apex via contact with two of the three gp120 protomers, possibly contributing to its reported preference for a quaternary epitope. Molecular modeling and isothermal titration calorimetry binding experiments with an engineered PG9 mutant suggest that, in addition to the N156 and N160 glycan interactions observed in crystal structures of PG9 with a scaffolded V1/V2 domain, PG9 makes secondary interactions with an N160 glycan from an adjacent gp120 protomer in the antibody-trimer complex. Together, these structural and biophysical findings should facilitate the design of HIV-1 immunogens that possess all elements of the quaternary PG9 epitope required to induce broadly neutralizing antibodies against this region. R ational immunogen design is an increasingly promising approach for development of an effective human immunodeficiency virus-1 (HIV-1) vaccine. The recent discovery of many new and potent broadly neutralizing antibodies (bnAbs) has helped define conserved sites of vulnerability on the HIV-1 envelope (Env) glycoprotein (gp) complex that mediates viral entry into cells (refs. 1-6 and reviewed in refs. 7-11). Passive immunization studies show that sterilizing immunity can be achieved if sufficient amounts of bnAbs are present before virus challenge in macaques (12-16). Hence, intensive efforts are ongoing to design immunogens capable of re-eliciting these types of bnAbs by vaccination.The major difficulty in mounting an effective antibody response against HIV-1 resides in the multiple evasion strategies that have evolved in Env. An error-prone reverse transcriptase drives a high degree of Env sequence diversity (17-19). The few conserved regions of Env are shielded by an extensive array of glycans (20-24) and are often occluded by more variable structures, such as the V1-V5 loops. However, because some HIV-1-infected individuals can develop bnAbs over the course of infection, these various evasion strategies are not insurmountable (1,(25)(26)(27). Although bnAbs do not seem to confer significant protection against disease progression in infected individuals (28, 29), their induction through vaccination might prevent the acquisition of infection. ...

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Section: Discussionmentioning

confidence: 89%

Section: Molecular Structure Revealed By Negative Stain Single Particmentioning

confidence: 94%

mentioning

confidence: 99%

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Asymmetric recognition of the HIV-1 trimer by broadly neutralizing antibody PG9

Julien

Lee

Cupo

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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230

303

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“…Even though one should be cautious in interpreting a peptide's conformational states and extrapolating back to the native functional protein, the potential relevance of the helical conformation adopted by MPERp N-terminal residues is emphasized by the structure of an antigenically near-native Env construct termed "SOSIP" gp140 (79). Although truncated at position 664, the recently solved crystal structure at 4.7 Å provides insights into the gp41 ectodomain organization in the context of cleaved, stabilized HIV-1 Env trimers (80).…”

Section: Figure 8 Recovered Responses After Vaccination With the Popmentioning

confidence: 99%

Structure and Immunogenicity of a Peptide Vaccine, Including the Complete HIV-1 gp41 2F5 Epitope

Serrano

Araujo

Apellániz

et al. 2014

Journal of Biological Chemistry

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Background: HIV-1 vaccines should elicit broadly neutralizing antibodies as the gp41 "membrane-proximal external region" targeting MAb2F5. Results: NMR disclosed unprecedented 2F5 peptide-epitope structures. Although overall conformation was preserved in different adjuvants, recovered antibodies after vaccination were functionally different. Conclusion: Membrane-inserted helical oligomers may encompass effective 2F5 peptide vaccines. Significance: Disclosing the structures that generate 2F5-like antibodies may guide future vaccine development.

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“…The glycans shields the envelope glycoproteins that can be studied better by deglycosylation [46]. The neutralizing ability of the antibodies targeted to specific epitopes is less efficient due to the carbohydrate shield without which the antibodies would neutralize better [47].…”

Section: Glycosylationmentioning

confidence: 99%

Conventional Vaccine to Prevent AIDS A Paradigm or A Paradox? A SWOT Analysis

Christdas¹

2016

HIV Curr Res

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AIDS as a disease afflicts mankind for more than 3 decades. HIV, the causative is not yet checked with a prophylactic vaccine. The plausibility of the prevention by conventional prophylactic strategies based on Jenner's conventional method of inducing memory response is reviewed here. In order to find the right direction towards achieving this goal, a SWOT analysis is represented here with a Venn diagram. Our increasing awareness on the viral genome and the antiviral treatment with the sensitive diagnostic tools strengthen the efforts, while the viral nature privileges its replenishment by hampering the host immune memory. Though antiviral drugs promise a notable degree of control, they render selection pressure favoring viral escapism with perking quasispecies or circulating recombinant forms (CRF). Few historical clinical trials teach us to frame new opportunities for the conduct of such trials that assures safety. The knowledge has exponentially increased on the host immune response, human genetics, retrovirology, drug development, gene delivery system to understand that HIV is the only pathogen that had the greatest consumption of our time and resources. The technological advancements add to our strength. However, the virus with its biological features proves its intrinsic competency for survival. Though the antiviral therapy confers some hope, the resistant forms pose a threat to their continuation. It emphasizes the need for prevention and suggests novel, unconventional prophylaxis. Treatment as a means of prevention and the use of immunomodulators to decrease the disease progression can help us to win the belligerence against AIDS.

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Partial Enzymatic Deglycosylation Preserves the Structure of Cleaved Recombinant HIV-1 Envelope Glycoprotein Trimers

Cited by 50 publications

References 83 publications

Asymmetric recognition of the HIV-1 trimer by broadly neutralizing antibody PG9

Asymmetric recognition of the HIV-1 trimer by broadly neutralizing antibody PG9

Structure and Immunogenicity of a Peptide Vaccine, Including the Complete HIV-1 gp41 2F5 Epitope

Conventional Vaccine to Prevent AIDS A Paradigm or A Paradox? A SWOT Analysis

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