HIV-1 Envelope and MPER Antibody Structures in Lipid Assemblies

Rantalainen, Kimmo; Berndsen, Zachary T.; Antanasijevic, Aleksandar; Schiffner, Torben; Zhang, Xi; Lee, Wen-Hsin; Torres, Jonathan L.; Zhang, Lei; Irimia, A.; Copps, Jeffrey; Zhou, Kenneth H.; Kwon, Young Do; Law, William H.; Schramm, Chaim A.; Verardi, Raffaello; Krebs, Shelly J.; Kwong, Peter D.; Doria‐Rose, Nicole A.; Wilson, Ian A.; Zwick, Michael B.; Yates, John R.; Schief, William R.; Ward, Andrew B.

doi:10.1016/j.celrep.2020.107583

Cited by 68 publications

(68 citation statements)

References 85 publications

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“…Several purification protocols have been developed that sustain membrane proteins in a native membrane-like environment (16)(17)(18)(19). In this study, we reconstituted Pdr5 in peptidiscs, which are short amphipathic bi-helical peptides (17) compatible with single-particle cryo-EM (20,21), after purification from S. cerevisiae cell membranes. This yielded homogenous particles readily visualised by electron cryo-microscopy ( Fig.…”

Section: Pdr5 Reconstituted In a Detergent-free System Can Be Imagedmentioning

confidence: 99%

Structure and efflux mechanism of the yeast pleiotropic drug resistance transporter Pdr5

Harris

Wagner

et al. 2021

Preprint

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Pdr5, a member of the extensive ABC transporter superfamily, is representative of a clinically relevant subgroup involved in pleiotropic drug resistance. Through the coupling of nucleotide hydrolysis with drug efflux, Pdr5 homologues enable pathogenic species to survive in the presence of chemically diverse antifungal agents. Our structural and functional results reveal details of an ATP-driven conformational cycle, which mechanically drives drug translocation through an amphipathic channel, and a clamping switch within a conserved linker loop that acts as a nucleotide sensor. One half of the transporter remains nearly invariant throughout the cycle, while its partner undergoes changes that are transmitted across inter-domain interfaces to support a peristaltic motion of the pumped molecule. The efflux model proposed here rationalises the pleiotropic impact of Pdr5 and opens avenues for the development of effective antifungal compounds.

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Section: Pdr5 Reconstituted In a Detergent-free System Can Be Imagedmentioning

confidence: 99%

Structure and efflux mechanism of the yeast pleiotropic drug resistance transporter Pdr5

Harris

Wagner

et al. 2021

Preprint

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“…With the aim to further characterize the functional aspect of the intermolecular conformations of Env, we investigated the effect of well-known HIV-1 broad neutralizing antibodies (bNAbs) on Env cluster formation. For this experiment, we selected three different antibodies recognizing separated regions of Env: PGT145, which recognizes the HIV-1 apex 28 ; B12, which binds to an overlapping region of gp120 with the site of CD4 attachment 29,30 ; and 10E8 which is directed against the membrane-proximal external region (MPER) 31 . Moreover, these antibodies show high (PGT145 and 10E8) to moderate (b12) ability to neutralize different HIV-1 isolates 32 .…”

Section: Resultsmentioning

confidence: 99%

“…B12, as opposed to CD4, is unable to bind the closed conformation of Env, although upon binding, b12 prevents reversion back to the closed state 29 , thus stabilizing an intermediate/open conformation 30 . Finally, 10E8 targets a quaternary epitope including lipid and MPER contacts 31 stabilizing an open conformation 25 .…”

Section: Resultsmentioning

confidence: 99%

Structure and Dynamics of HIV-1 Env Trimers on Native Virions Engaged in Living T Cells

Carlón-Andrés

Malinauskas

Padilla‐Parra

2020

Preprint

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The HIV-1 envelope glycoprotein (Env) mediates viral entry into the host cell. Although the highly dynamic nature of Env intramolecular conformations has been shown with single molecule spectroscopy in vitro, the bona fide Env intra- and intermolecular mechanics when engaged in live T cells remains unknown. We used both, two photon fast fluorescence lifetime imaging detection of single-molecule Förster Resonance Energy Transfer and single molecule photobleaching to reveal transitions between intramolecular and intermolecular conformations that mediate Env clustering. Furthermore, we show that three broad neutralizing anti-Env antibodies directed to different epitopes destabilise Env intramolecular dynamics and their clusters when engaged to living T cells. Importantly, our results show that Env clustering is a common conformation across different HIV-1 Env strains, which depends on efficient virus maturation, and that is disrupted upon binding of Env to CD4 or to neutralizing antibodies. Thus, this study illuminates how different intramolecular conformations and clusters of Env mediate HIV-1 Env–T cell interactions in real time and therefore might control immune evasion.

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“…The open pore state of bacterial Tc toxin complex TcdA1, which could not be sufficiently stabilized in Tween-20 detergent micelles or liposomes [143,144], could be resolved in high detail embedded in nanodiscs [145], whereas the corresponding crystal structure could only reveal a closed pre-pore state [143]. Likewise, when characterized in nanodiscs, aforementioned HIV envelope protein Env showed substantial differences in its arrangement and orientation to the membrane surface, compared to micellar and bicellar environments [146].…”

Section: Bicelles and Nanodiscsmentioning

confidence: 99%

Fake It ‘Till You Make It—The Pursuit of Suitable Membrane Mimetics for Membrane Protein Biophysics

Thoma

Burmann

2020

IJMS

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Membrane proteins evolved to reside in the hydrophobic lipid bilayers of cellular membranes. Therefore, membrane proteins bridge the different aqueous compartments separated by the membrane, and furthermore, dynamically interact with their surrounding lipid environment. The latter not only stabilizes membrane proteins, but directly impacts their folding, structure and function. In order to be characterized with biophysical and structural biological methods, membrane proteins are typically extracted and subsequently purified from their native lipid environment. This approach requires that lipid membranes are replaced by suitable surrogates, which ideally closely mimic the native bilayer, in order to maintain the membrane proteins structural and functional integrity. In this review, we survey the currently available membrane mimetic environments ranging from detergent micelles to bicelles, nanodiscs, lipidic-cubic phase (LCP), liposomes, and polymersomes. We discuss their respective advantages and disadvantages as well as their suitability for downstream biophysical and structural characterization. Finally, we take a look at ongoing methodological developments, which aim for direct in-situ characterization of membrane proteins within native membranes instead of relying on membrane mimetics.

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HIV-1 Envelope and MPER Antibody Structures in Lipid Assemblies

Cited by 68 publications

References 85 publications

Structure and efflux mechanism of the yeast pleiotropic drug resistance transporter Pdr5

Structure and efflux mechanism of the yeast pleiotropic drug resistance transporter Pdr5

Structure and Dynamics of HIV-1 Env Trimers on Native Virions Engaged in Living T Cells

Fake It ‘Till You Make It—The Pursuit of Suitable Membrane Mimetics for Membrane Protein Biophysics

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