The Atomic Structure of the HIV-1 gp41 Transmembrane Domain and Its Connection to the Immunogenic Membrane-proximal External Region

Apellániz, Beatriz; Rujas, Edurne; Serrano, Susana; Morante, Koldo; Tsumoto, Kouhei; Caaveiro, José M. M.; Jiménez, María Ángeles Sánchez; Nieva, José L.

doi:10.1074/jbc.m115.644351

Cited by 40 publications

(72 citation statements)

References 82 publications

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“…Excluding the Lys solubility tags and Asp664, the resulting structure ensembles were well defined, as indicated by the small RSMD values for the backbone atoms of residues 665–690 (0.7 ± 0.2 Å in HFIP, and 0.7 ± 0.2 Å in DPC; see Supplementary Table 1). Confirming the structural motif inferred from the previous reconstitution process13, these helical structures exhibited a kink at position 671 NW 672 . The variability on the orientation of the two helices within each structural ensemble and the prominent changes in the direction of the helix axis at this position in two selected structures can be appreciated in Fig.…”

Section: Resultssupporting

confidence: 83%

“…1a), termed “MPER-N-TMD” in our recent work13, revised models which assumed that the interfacial MPER helix bends at position Lys683 to promote the insertion of the TMD perpendicular to the plane of the membrane171819202122. We hypothesized that peptides derived from the region MPER-N-TMD might function as a helical scaffold to increase the affinity of antibodies targeting the C-terminal subregion of the MPER13. We therefore designed the peptide 10E8ep, KKKK - 664 D K WA S LW- NW-F DI T NW LW YI K LFIMIVG 690 - KKKKK , to cover the entire length of the H1-H2 helices (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In this study, we have elucidated the structure of the 10E8 Fab in complex with a peptide antigen whose affinity has been optimized by the addition of native residues belonging to the gp41 transmembrane domain (TMD)13. This peptide has been termed 10E8ep.…”

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confidence: 99%

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Structural basis for broad neutralization of HIV-1 through the molecular recognition of 10E8 helical epitope at the membrane interface

Rujas

Caaveiro

Partida-Hanon

et al. 2016

Sci Rep

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The mechanism by which the HIV-1 MPER epitope is recognized by the potent neutralizing antibody 10E8 at membrane interfaces remains poorly understood. To solve this problem, we have optimized a 10E8 peptide epitope and analyzed the structure and binding activities of the antibody in membrane and membrane-like environments. The X-ray crystal structure of the Fab-peptide complex in detergents revealed for the first time that the epitope of 10E8 comprises a continuous helix spanning the gp41 MPER/transmembrane domain junction (MPER-N-TMD; Env residues 671–687). The MPER-N-TMD helix projects beyond the tip of the heavy-chain complementarity determining region 3 loop, indicating that the antibody sits parallel to the plane of the membrane in binding the native epitope. Biophysical, biochemical and mutational analyses demonstrated that strengthening the affinity of 10E8 for the TMD helix in a membrane environment, correlated with its neutralizing potency. Our research clarifies the molecular mechanisms underlying broad neutralization of HIV-1 by 10E8, and the structure of its natural epitope. The conclusions of our research will guide future vaccine-design strategies targeting MPER.

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Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

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Structural basis for broad neutralization of HIV-1 through the molecular recognition of 10E8 helical epitope at the membrane interface

Rujas

Caaveiro

Partida-Hanon

et al. 2016

Sci Rep

Self Cite

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“…Similarly, the membrane-proximal external region (MPER) and MSD of HIV-1 Env have been shown to be crucial for promoting fusogenicity. Disruption of hinge regions or the tryptophan-rich sequence in the MPER of HIV-1 Env, as well as polar residues within the MSD, all result in nonfunctional glycoproteins that are fusogenically inactive (24)(25)(26)(27)(28)(29). The potential role for the MSDs of MLV and HIV-1 Env in potentiating molecular rearrangements necessary for fusogenic activity would be analogous to the MSDs of voltage-gated ion channels and G-protein-coupled receptors (GPCRs).…”

mentioning

confidence: 99%

Characterizing the Murine Leukemia Virus Envelope Glycoprotein Membrane-Spanning Domain for Its Roles in Interface Alignment and Fusogenicity

Salamango

Johnson

2015

J Virol

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The membrane-proximal region of murine leukemia virus envelope (Env) is a critical modulator of its functionality. We have previously shown that the insertion of one amino acid (؉1 leucine) within the membrane-spanning domain (MSD) abolished protein functionality in infectivity assays. However, functionality could be restored to this ؉1 leucine mutant by either inserting two additional amino acids (؉3 leucine) or by deleting the cytoplasmic tail domain (CTD) in the ؉1 leucine background. We inferred that the ectodomain and CTD have protein interfaces that have to be in alignment for Env to be functional. Here, we made single residue deletions to the Env mutant with the ؉1 leucine insertion to restore the interface alignment (gain of functionality) and therefore define the boundaries of the two interfaces. We identified the glycine-proline pairs near the N terminus (positions 147 and 148) and the C terminus (positions 159 and 160) of the MSD as being the boundaries of the two interfaces. Deletions between these pairs restored function, but deletions outside of them did not. In addition, the vast majority of the single residue deletions regained function if the CTD was deleted. The exceptions were four hydroxyl-containing amino acid residues (T139, T140, S143, and T144) that reside in the ectodomain interface and the proline at position 148, which were all indispensable for functionality. We hypothesize that the hydroxyl-containing residues at positions T139 and S143 could be a driving force for stabilizing the ectodomain interface through formation of a hydrogen-bonding network. IMPORTANCEThe membrane-proximal external region (MPER) and membrane-spanning domains (MSDs) of viral glycoproteins have been shown to be critical for regulating glycoprotein fusogenicity. However, the roles of these two domains are poorly understood. We report here that point deletions and insertions within the MPER or MSD result in functionally inactive proteins. However, when the C-terminal tail domain (CTD) is deleted, the majority of the proteins remain functional. The only residues that were found to be critical for function regardless of the CTD were four hydroxyl-containing amino acids located at the C terminus of the MPER (T139 and T140) and at the N terminus of the MSD (S143 and T144) and a proline near the beginning of the MSD (P148). We demonstrate that hydrogen-bonding at positions T139 and S143 is critical for protein function. Our findings provide novel insights into the role of the MPER in regulating fusogenic activity of viral glycoproteins. E nveloped viruses require membrane-spanning cell surface glycoproteins to coordinate fusion between viral and host cell membranes. Retroviral envelope (Env) glycoproteins are produced as precursors that undergo trimerization in the endoplasmic reticulum (1-3). Subsequently, the precursor trimer is cleaved into two subdomains: the receptor-binding surface domain (SU) and the fusion-promoting transmembrane domain (TM). This cleavage process is mediated by a host-cell furin, or f...

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“…Alde batetik, itu-zelularen mintzean txertatuko da gp41 azpiunitatearen amino muturrean kokatzen den fusio peptidoa (FP); beste aldetik, mintza desegonkortzen lagundu lezake eskualde jakin batek, alegia mintza zeharkatzen duen domeinuaren [7][8][9][10].…”

Section: Gib Birusaunclassified

GIBaren aurkako txertoaren bila

Torralba¹,

Goikoetxea²,

Apellániz

2017

EKAIA

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Laburpena: GIB birusa, giza historian zehar gertatu den pandemiarik hilgarrienetariko baten eragilea da, HIESa alegia. Nahiz eta azken urteetan HAART terapia antirretrobiralari esker heriotza tasa asko murriztu den, infekzio berrien kopurua etengabe emendatzen ari da. Hori dela eta, badirudi pandemiari azkenik amaiera eman ahal izateko txerto prebentibo baten garapena izan daitekeela jokaera eraginkorrena. Hala ere, birusak garatu dituen estrategia natural desberdinek eta immunizazioz sortu nahi diren antigorputz neutralizatzaileen ezaugarri bereziek oso zaila egiten dute infekziotik babestuko lukeen txertoaren garapena. RV144 entsegu klinikoaren eraginkortasun apalek eta espektro zabaleko antigorputz neutralizatzaileekin immunizazio pasiboz lorturiko emaitzek iradokitzen dute hurbilago gaudela txerto eraginkor baten garapena lortzetik. Orain arte hainbat txerto mota erabili diren arren, emaitza onik ez da lortu. Hala ere, sakon aztertzen ari da bai immunogeno naturalen egitura, bai immunogenoak ezagutzen dituzten antigorputzen egitura, eta badirudi, besteak beste, ikerketa horiek bide berriak irekiko dituztela txerto berrien diseinuan. Hitz gakoak: GIB, txertoa, antigorputz neutralizatzaileak, Env fusio-glukoproteina.Abstract: HIV is the causative agent of one of the most lethal pandemics in human history, AIDS. Although in recent years HAART therapy has considerably reduced death rate, new infections keep arising. Therefore, a preventive vaccine remains the most promising tool to end the pandemic. However, the virus has developed many natural strategies to evade the immune system and, moreover, the special characteristics of the neutralizing antibodies that a vaccine aims to elicit, makes vaccine design extremely challenging. The efficiency of the RV144 clinical trial and the promising results obtained by passive immunization with broadly neutralizing antibodies suggest that the objective of obtaining an effective vaccine is closer. Until now, a myriad of vaccine strategies have been attempted but the expected results have not been yet ob-EKAIA 32.indd 7 EKAIA 32.indd 7

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The Atomic Structure of the HIV-1 gp41 Transmembrane Domain and Its Connection to the Immunogenic Membrane-proximal External Region

Cited by 40 publications

References 82 publications

Structural basis for broad neutralization of HIV-1 through the molecular recognition of 10E8 helical epitope at the membrane interface

Structural basis for broad neutralization of HIV-1 through the molecular recognition of 10E8 helical epitope at the membrane interface

Characterizing the Murine Leukemia Virus Envelope Glycoprotein Membrane-Spanning Domain for Its Roles in Interface Alignment and Fusogenicity

GIBaren aurkako txertoaren bila

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