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A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus

Aromatic proton shifts during dilution of PBS-buffered CLR01.Upfield shifts of the inner aromatic protons (2-H, 3-H, 13-H, 14-H) document their mutual inclusion inside the cavity of the other host molecule, leading to specific anisotropic shielding exclusively in the dimer. Thus, the NMR shift changes prove formation of weak dimers (Ka ∼60 M−1) and exclude unspecific aggregation, which would manifest as smaller chemical shift changes evenly distributed among the CH protons of CLR01.DOI:http://dx.doi.org/10.7554/eLife.05397.024
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fig7s1: Aromatic proton shifts during dilution of PBS-buffered CLR01.Upfield shifts of the inner aromatic protons (2-H, 3-H, 13-H, 14-H) document their mutual inclusion inside the cavity of the other host molecule, leading to specific anisotropic shielding exclusively in the dimer. Thus, the NMR shift changes prove formation of weak dimers (Ka ∼60 M−1) and exclude unspecific aggregation, which would manifest as smaller chemical shift changes evenly distributed among the CH protons of CLR01.DOI:http://dx.doi.org/10.7554/eLife.05397.024

Mentions: To investigate whether CLR01 even forms colloidal aggregates, we performed various tests for CLR01 aggregation using the same buffers and CLR01 concentrations as employed in our biological experiments described above (Figures 2A–C, 3A–C, 5A). NMR dilution titrations provided experimental evidence for a very weak CLR01 dimer formation (Ka = ∼60 M−1) in PBS buffer (10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl, pH 7.4), with ∼5% CLR01 dimers present when the CLR01 concentration was 500 µM (Figure 7A, Figure 7—figure supplement 1). This observation is consistent with previously published data (Dutt et al., 2013). Dimerization was totally absent in HEPES buffer (25 mM HEPES, 150 mM KOAc, 10 mM Mg(OAc)2, pH 7.4; Figure 7B, Figure 7—figure supplement 2). Thus, CLR01 is predominantly monomeric.10.7554/eLife.05397.023Figure 7.CLR01 does not form colloid or micelle aggregates.


A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

Aromatic proton shifts during dilution of PBS-buffered CLR01.Upfield shifts of the inner aromatic protons (2-H, 3-H, 13-H, 14-H) document their mutual inclusion inside the cavity of the other host molecule, leading to specific anisotropic shielding exclusively in the dimer. Thus, the NMR shift changes prove formation of weak dimers (Ka ∼60 M−1) and exclude unspecific aggregation, which would manifest as smaller chemical shift changes evenly distributed among the CH protons of CLR01.DOI:http://dx.doi.org/10.7554/eLife.05397.024
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4536748&req=5

fig7s1: Aromatic proton shifts during dilution of PBS-buffered CLR01.Upfield shifts of the inner aromatic protons (2-H, 3-H, 13-H, 14-H) document their mutual inclusion inside the cavity of the other host molecule, leading to specific anisotropic shielding exclusively in the dimer. Thus, the NMR shift changes prove formation of weak dimers (Ka ∼60 M−1) and exclude unspecific aggregation, which would manifest as smaller chemical shift changes evenly distributed among the CH protons of CLR01.DOI:http://dx.doi.org/10.7554/eLife.05397.024
Mentions: To investigate whether CLR01 even forms colloidal aggregates, we performed various tests for CLR01 aggregation using the same buffers and CLR01 concentrations as employed in our biological experiments described above (Figures 2A–C, 3A–C, 5A). NMR dilution titrations provided experimental evidence for a very weak CLR01 dimer formation (Ka = ∼60 M−1) in PBS buffer (10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl, pH 7.4), with ∼5% CLR01 dimers present when the CLR01 concentration was 500 µM (Figure 7A, Figure 7—figure supplement 1). This observation is consistent with previously published data (Dutt et al., 2013). Dimerization was totally absent in HEPES buffer (25 mM HEPES, 150 mM KOAc, 10 mM Mg(OAc)2, pH 7.4; Figure 7B, Figure 7—figure supplement 2). Thus, CLR01 is predominantly monomeric.10.7554/eLife.05397.023Figure 7.CLR01 does not form colloid or micelle aggregates.

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus