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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

CLR01 does not displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils.Preformed SEVI, PAP85-120, or SEM1(45-107) fibrils (5 µM monomer) were preincubated with ThT (25 µM) for 30 min at room temperature. Buffer (PBS), CLR01 (250 µM), or a known competitor of ThT binding, BTA-1 (250 µM) were then added and incubated for 10 min at room temperature. ThT displacement was then assessed by fluorescence measurements. Values represent means ±SEM (n = 3). A one-way ANOVA with the post hoc Dunnett's multiple comparisons test was used to compare the buffer control to the CLR01 or BTA-1 condition (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.005
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fig2s1: CLR01 does not displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils.Preformed SEVI, PAP85-120, or SEM1(45-107) fibrils (5 µM monomer) were preincubated with ThT (25 µM) for 30 min at room temperature. Buffer (PBS), CLR01 (250 µM), or a known competitor of ThT binding, BTA-1 (250 µM) were then added and incubated for 10 min at room temperature. ThT displacement was then assessed by fluorescence measurements. Values represent means ±SEM (n = 3). A one-way ANOVA with the post hoc Dunnett's multiple comparisons test was used to compare the buffer control to the CLR01 or BTA-1 condition (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.005

Mentions: We examined the possibility that CLR01 might simply displace ThT from fibrils by employing a ThT displacement assay (Lockhart et al., 2005). Thus, preformed SEVI, PAP85-120, or SEM1(45-107) fibrils were preincubated with ThT. Buffer, or an excess of CLR01 or a known competitor of ThT binding, BTA-1, were then added and ThT displacement was assessed by fluorescence measurements (Lockhart et al., 2005). ThT fluorescence decreased drastically in the presence of BTA-1, confirming its ability to displace ThT from fibrils (Lockhart et al., 2005), whereas CLR01 and buffer had no effect (Figure 2—figure supplement 1). These findings suggest that CLR01 does not simply displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils. Thus, any reduction in ThT fluorescence caused by CLR01 can be attributed to an inhibition of fibril assembly.


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)

CLR01 does not displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils.Preformed SEVI, PAP85-120, or SEM1(45-107) fibrils (5 µM monomer) were preincubated with ThT (25 µM) for 30 min at room temperature. Buffer (PBS), CLR01 (250 µM), or a known competitor of ThT binding, BTA-1 (250 µM) were then added and incubated for 10 min at room temperature. ThT displacement was then assessed by fluorescence measurements. Values represent means ±SEM (n = 3). A one-way ANOVA with the post hoc Dunnett's multiple comparisons test was used to compare the buffer control to the CLR01 or BTA-1 condition (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.005
© Copyright Policy
Related In: Results  -  Collection

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

fig2s1: CLR01 does not displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils.Preformed SEVI, PAP85-120, or SEM1(45-107) fibrils (5 µM monomer) were preincubated with ThT (25 µM) for 30 min at room temperature. Buffer (PBS), CLR01 (250 µM), or a known competitor of ThT binding, BTA-1 (250 µM) were then added and incubated for 10 min at room temperature. ThT displacement was then assessed by fluorescence measurements. Values represent means ±SEM (n = 3). A one-way ANOVA with the post hoc Dunnett's multiple comparisons test was used to compare the buffer control to the CLR01 or BTA-1 condition (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.005
Mentions: We examined the possibility that CLR01 might simply displace ThT from fibrils by employing a ThT displacement assay (Lockhart et al., 2005). Thus, preformed SEVI, PAP85-120, or SEM1(45-107) fibrils were preincubated with ThT. Buffer, or an excess of CLR01 or a known competitor of ThT binding, BTA-1, were then added and ThT displacement was assessed by fluorescence measurements (Lockhart et al., 2005). ThT fluorescence decreased drastically in the presence of BTA-1, confirming its ability to displace ThT from fibrils (Lockhart et al., 2005), whereas CLR01 and buffer had no effect (Figure 2—figure supplement 1). These findings suggest that CLR01 does not simply displace ThT from SEVI, PAP85-120, or SEM1(45-107) fibrils. Thus, any reduction in ThT fluorescence caused by CLR01 can be attributed to an inhibition of fibril assembly.

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