Neutralizing epitopes in the membrane-proximal external region of HIV-1 gp41 Are influenced by the transmembrane domain and the plasma membrane

Marinieve Montero, Naveed Gulzar, Kristina Ana Klaric, Jason E. Donald, Christa Lepik, Sampson Wu, Sue Tsai, Jean Philippe Julien, Ann Hessell, Shixia Wang, Shan Lu, Dennis R. Burton, Emil F. Pai, William F. DeGrado, Jamie K. Scott

    Research output: Contribution to journalArticle

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    Abstract

    Failure to elicit broadly neutralizing (bNt) antibodies (Abs) against the membrane-proximal external region of HIV-1 gp41 (MPER) reflects the difficulty of mimicking its neutralization-competent structure (NCS). Here, we analyzed MPER antigenicity in the context of the plasma membrane and identified a role for the gp41 transmembrane domain (TM) in exposing the epitopes of three bNt monoclonal Abs (MAbs) (2F5, 4E10, and Z13e1). We transiently expressed DNA constructs encoding gp41 ectodomain fragments fused to either the TM of the platelet-derived growth factor receptor (PDGFR) or the gp41 TM and cytoplasmic tail domain (CT). Constructs encoding the MPER tethered to the gp41 TM followed by a 27-residue CT fragment (MPER-TM1) produced optimal MAb binding. Critical binding residues for the three Nt MAbs were identified using a panel of 24 MPER-TM1 mutants bearing single amino acid substitutions in the MPER; many were previously shown to affect MAb-mediated viral neutralization. Moreover, non-Nt mutants of MAbs 2F5 and 4E10 exhibited a reduction in binding to MPER-TM1 and yet maintained binding to synthetic MPER peptides, indicating that MPER-TM1 better approximates the MPER NCS than peptides. Replacement of the gp41 TM and CT of MPER-TM1 with the PDGFR TM reduced binding by MAb 4E10, but not 2F5, indicating that the gp41 TM plays a pivotal role in orienting the 4E10 epitope, and more globally, in affecting MPER exposure.

    Original languageEnglish (US)
    Pages (from-to)2930-2941
    Number of pages12
    JournalJournal of Virology
    Volume86
    Issue number6
    DOIs
    StatePublished - Mar 2012

    Fingerprint

    Human immunodeficiency virus 1
    neutralization
    epitopes
    HIV-1
    Epitopes
    plasma membrane
    Cell Membrane
    Membranes
    Platelet-Derived Growth Factor Receptors
    tail
    Neutralizing Antibodies
    neutralizing antibodies
    artificial membranes
    peptides
    mutants
    Peptides
    amino acid substitution
    Amino Acid Substitution
    monoclonal antibodies
    Monoclonal Antibodies

    ASJC Scopus subject areas

    • Immunology
    • Virology

    Cite this

    Neutralizing epitopes in the membrane-proximal external region of HIV-1 gp41 Are influenced by the transmembrane domain and the plasma membrane. / Montero, Marinieve; Gulzar, Naveed; Klaric, Kristina Ana; Donald, Jason E.; Lepik, Christa; Wu, Sampson; Tsai, Sue; Julien, Jean Philippe; Hessell, Ann; Wang, Shixia; Lu, Shan; Burton, Dennis R.; Pai, Emil F.; DeGrado, William F.; Scott, Jamie K.

    In: Journal of Virology, Vol. 86, No. 6, 03.2012, p. 2930-2941.

    Research output: Contribution to journalArticle

    Montero, M, Gulzar, N, Klaric, KA, Donald, JE, Lepik, C, Wu, S, Tsai, S, Julien, JP, Hessell, A, Wang, S, Lu, S, Burton, DR, Pai, EF, DeGrado, WF & Scott, JK 2012, 'Neutralizing epitopes in the membrane-proximal external region of HIV-1 gp41 Are influenced by the transmembrane domain and the plasma membrane', Journal of Virology, vol. 86, no. 6, pp. 2930-2941. https://doi.org/10.1128/JVI.06349-11
    Montero, Marinieve ; Gulzar, Naveed ; Klaric, Kristina Ana ; Donald, Jason E. ; Lepik, Christa ; Wu, Sampson ; Tsai, Sue ; Julien, Jean Philippe ; Hessell, Ann ; Wang, Shixia ; Lu, Shan ; Burton, Dennis R. ; Pai, Emil F. ; DeGrado, William F. ; Scott, Jamie K. / Neutralizing epitopes in the membrane-proximal external region of HIV-1 gp41 Are influenced by the transmembrane domain and the plasma membrane. In: Journal of Virology. 2012 ; Vol. 86, No. 6. pp. 2930-2941.
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    abstract = "Failure to elicit broadly neutralizing (bNt) antibodies (Abs) against the membrane-proximal external region of HIV-1 gp41 (MPER) reflects the difficulty of mimicking its neutralization-competent structure (NCS). Here, we analyzed MPER antigenicity in the context of the plasma membrane and identified a role for the gp41 transmembrane domain (TM) in exposing the epitopes of three bNt monoclonal Abs (MAbs) (2F5, 4E10, and Z13e1). We transiently expressed DNA constructs encoding gp41 ectodomain fragments fused to either the TM of the platelet-derived growth factor receptor (PDGFR) or the gp41 TM and cytoplasmic tail domain (CT). Constructs encoding the MPER tethered to the gp41 TM followed by a 27-residue CT fragment (MPER-TM1) produced optimal MAb binding. Critical binding residues for the three Nt MAbs were identified using a panel of 24 MPER-TM1 mutants bearing single amino acid substitutions in the MPER; many were previously shown to affect MAb-mediated viral neutralization. Moreover, non-Nt mutants of MAbs 2F5 and 4E10 exhibited a reduction in binding to MPER-TM1 and yet maintained binding to synthetic MPER peptides, indicating that MPER-TM1 better approximates the MPER NCS than peptides. Replacement of the gp41 TM and CT of MPER-TM1 with the PDGFR TM reduced binding by MAb 4E10, but not 2F5, indicating that the gp41 TM plays a pivotal role in orienting the 4E10 epitope, and more globally, in affecting MPER exposure.",
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    AU - Montero, Marinieve

    AU - Gulzar, Naveed

    AU - Klaric, Kristina Ana

    AU - Donald, Jason E.

    AU - Lepik, Christa

    AU - Wu, Sampson

    AU - Tsai, Sue

    AU - Julien, Jean Philippe

    AU - Hessell, Ann

    AU - Wang, Shixia

    AU - Lu, Shan

    AU - Burton, Dennis R.

    AU - Pai, Emil F.

    AU - DeGrado, William F.

    AU - Scott, Jamie K.

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