Membrane-transferring Sequences of the HIV-1 Gp41 Ectodomain Assemble into an Immunogenic Complex

Lorizate, Maier; Gómara, María J.; Torre, Beatriz G. de la; Andreu, David; Nieva, José L.

doi:10.1016/j.jmb.2006.04.056

Cited by 40 publications

(58 citation statements)

References 50 publications

(65 reference statements)

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“…A more recent report from Lorizate et al provides evidence that the 2F5 epitope may be supported by the N-terminal region of gp41 (131). Those authors hypothesized that, in the native or prefusion structure of the spike, the FP and the MPER are located at the same end of the ectodomain and that this arrange-ment helps to maintain the gp41 structure recognized by MAb 2F5 (131).…”

Section: Mab 2f5 and Its Epitopementioning

confidence: 99%

The Membrane-Proximal External Region of the Human Immunodeficiency Virus Type 1 Envelope: Dominant Site of Antibody Neutralization and Target for Vaccine Design

Montero¹,

Houten

Wang³

et al. 2008

Microbiol Mol Biol Rev

233

194

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SUMMARY Enormous efforts have been made to produce a protective vaccine against human immunodeficiency virus type 1; there has been little success. However, the identification of broadly neutralizing antibodies against epitopes on the highly conserved membrane-proximal external region (MPER) of the gp41 envelope protein has delineated this region as an attractive vaccine target. Furthermore, emerging structural information on the MPER has provided vaccine designers with new insights for building relevant immunogens. This review describes the current state of the field regarding (i) the structure and function of the gp41 MPER; (ii) the structure and binding mechanisms of the broadly neutralizing antibodies 2F5, 4E10, and Z13; and (iii) the development of an MPER-targeting vaccine. In addition, emerging approaches to vaccine design are presented.

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Section: Mab 2f5 and Its Epitopementioning

confidence: 99%

The Membrane-Proximal External Region of the Human Immunodeficiency Virus Type 1 Envelope: Dominant Site of Antibody Neutralization and Target for Vaccine Design

Montero¹,

Houten

Wang³

et al. 2008

Microbiol Mol Biol Rev

233

194

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“…A triglutamic acid tag was attached to the N′ peptide (EEE, acidic, pK a = 3), to adjust the pI of each peptide to approximately 5 ( Figure 3C, D). The solubility tags were added to the side of the peptide that contained the gp41 core segment (27) to prevent interference with possible interactions between the S528-Q540 and W666-N677 segments which are the major focus of this study. Clearly, the solubility tags altered the charge versus pH profile of the peptides (Figure 3), and we found that at pH 3.2 N29E3-R3C31 and N42E3-R3C43 complexes exhibited solubilities of up to 2 and 1.5 mM, respectively ( Table 2).…”

Section: Addition Of Solubility Tagsmentioning

confidence: 99%

The Membrane Proximal External Region of the HIV-1 Envelope Glycoprotein gp41 Contributes to the Stabilization of the Six-Helix Bundle Formed with a Matching N′ Peptide

et al. 2008

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The HIV-1 envelope glycoprotein gp41 undergoes a sequence of extensive conformational changes while participating in the fusion of the virus with the host cell. Since the discovery of its postfusion conformation, the structure and function of the protease-resistant six-helix bundle (6-HB) have been the subject of extensive investigation. In this work, we describe additional determinants (S528-Q540 and W666-N677) in the fusion peptide proximal region (FP-PR) and the membrane proximal external region (MPER) that stabilize the six-helix bundle and are involved in the interaction of T-20 (FUZEON, an anti-HIV-1 fusion inhibitor drug) with the gp41 FP-PR. Circular dichroism and sedimentation equilibrium measurements indicate that the 1:1 mixture of N′ and C′ peptides comprising residues A541-T569 and I635-K665 from the gp41 first and second helical repeats, HR1 and HR2, respectively, fail to form a stable six-helix bundle. Triglutamic acid and triarginine tags were added to these N′ and C′ peptides, respectively, at the termini distant from the FP-PR and the MPER to alter their pI and increase their solubility at pH 3.5. The tagged HR1 and HR2 peptides were elongated by addition of residues S528-Q540 from the FP-PR and residues W666-N677 from the MPER, respectively. A 1:1 complex of the elongated peptides formed a stable six-helix bundle which melted at 60 °C. These results underscore the importance of a detailed high-resolution characterization of MPER interactions, the results of which may improve our understanding of the structure-function relationship of gp41 and its role in HIV-1 fusion.The human immunodeficiency virus type 1 (HIV-1) 1 envelope protein is initially produced as a precursor glycoprotein gp160, which is proteolytically cleaved into two subunits. The resulting surface subunit (gp120) and transmembrane subunit (gp41) remain noncovalently associated and oligomerize as trimers on the surface of the virion. The transmembrane subunit gp41 mediates fusion between HIV-1 and its target cells (1). After the penetration of the gp41 † This study was supported by NIH Grants GM53329 (J.A.) and GM22086 (F. * To whom correspondence should be addressed. E-mail: Jacob.Anglister@weizmann.ac.il. . ‡ Weizmann Institute of Science. § College of Staten Island of the City University of New York.1 Abbreviations: CD, circular dichroism; DIEA, diisopropylethylamine; EDTA, ethylenediaminetetraacetic acid; EEE, Glu-Glu-Glu tag; FP-PR, fusion peptide proximal region; gp41, HIV-1 glycoprotein 41; gp120, HIV-1 glycoprotein 120; HIV-1, human immunodeficiency virus type 1; HPLC, high-pressure liquid chromatography; HR1, HIV-1 gp41 N′ heptad repeat 1; HR2, HIV-1 gp41 C′ heptad repeat 2; IPTG, isopropyl β-D-1-thiogalactopyranoside; K d , dissociation constant; MPER, membrane proximal external region; MALDI, matrixassisted laser desorption ionization; NMR, nuclear magnetic resonance; RRR, Arg-Arg-Arg tag; SIV, simian immunodeficiency virus; T-20, HIV fusion inhibitor corresponding to gp41 residues Y638-F673; TFA, trifluo-roacet...

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“…A similar study was not implemented for 2F5, because the case for 2F5-lipid interactions is less clear, its very occurrence a topic of current debate (21,27,31). Moreover, substitutions of hydrophobic residues at the apex of the 2F5 CDRH3 have been shown to affect antigen binding (32), which may relate to the proposed ability of 2F5 to form contacts with another region of Env on intact trimers (i.e., a region near the fusion peptide of gp41) (11,33).…”

mentioning

confidence: 99%

Aromatic residues at the edge of the antibody combining site facilitate viral glycoprotein recognition through membrane interactions

Scherer

Leaman

Zwick

et al. 2010

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

107

118

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The broadly neutralizing anti-HIV antibody 4E10 recognizes an epitope very close to the virus membrane on the glycoprotein gp41. It was previously shown that epitope recognition improves in a membrane context and that 4E10 binds directly, albeit weakly, to lipids. Furthermore, a crystal structure of Fab 4E10 complexed to an epitope peptide revealed that the centrally placed, protruding H3 loop of the antibody heavy chain does not form peptide contacts. To investigate the hypothesis that the H3 loop apex might interact with the viral membrane, two Trp residues in this region were substituted separately or in combination with either Ala or Asp by site-directed mutagenesis. The resultant IgG variants exhibited similar affinities for an epitope peptide as WT 4E10 but lower apparent affinities for both viral membrane mimetic liposomes and Env(−) virus. Variants also exhibited lower apparent affinities for Env(+) virions and failed to significantly neutralize a number of 4E10-sensitive viruses. For the extremely sensitive HXB2 virus, variants did neutralize, but at 37-to >250-fold lower titers than WT 4E10, with Asp substitutions exerting a greater effect on neutralization potency than Ala substitutions. Because reductions in lipid binding reflect trends in neutralization potency, we conclude that Trp residues in the antibody H3 loop enable membrane proximal epitope recognition through favorable lipid interactions. The requirement for lipophilic residues such as Trp adjacent to the antigen binding site may explain difficulties in eliciting 4E10-like neutralizing antibody responses by immunization and helps define a unique motif for antibody recognition of membrane proximal antigens.4E10 | gp41 | HIV | neutralizing antibody | tryptophan

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Membrane-transferring Sequences of the HIV-1 Gp41 Ectodomain Assemble into an Immunogenic Complex

Cited by 40 publications

References 50 publications

The Membrane-Proximal External Region of the Human Immunodeficiency Virus Type 1 Envelope: Dominant Site of Antibody Neutralization and Target for Vaccine Design

The Membrane-Proximal External Region of the Human Immunodeficiency Virus Type 1 Envelope: Dominant Site of Antibody Neutralization and Target for Vaccine Design

The Membrane Proximal External Region of the HIV-1 Envelope Glycoprotein gp41 Contributes to the Stabilization of the Six-Helix Bundle Formed with a Matching N′ Peptide

Aromatic residues at the edge of the antibody combining site facilitate viral glycoprotein recognition through membrane interactions

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