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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

BSA or preclearing CLR01 has no effect on the ability of CLR01 to inhibit formation of seminal amyloid fibrils.PAP248-286 (1 mM), PAP85-120 (1 mM), or SEM1(45-107) (500 µM) was incubated with CLR01 (1 mM) or buffer in the presence or absence of BSA (10 mg/ml) and agitated at 1400 rpm at 37°C for 68 hr. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibrillization was assessed via ThT fluorescence. 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 alone control to the other conditions (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.008
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fig2s4: BSA or preclearing CLR01 has no effect on the ability of CLR01 to inhibit formation of seminal amyloid fibrils.PAP248-286 (1 mM), PAP85-120 (1 mM), or SEM1(45-107) (500 µM) was incubated with CLR01 (1 mM) or buffer in the presence or absence of BSA (10 mg/ml) and agitated at 1400 rpm at 37°C for 68 hr. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibrillization was assessed via ThT fluorescence. 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 alone control to the other conditions (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.008

Mentions: Some small molecules must form higher order colloidal aggregates to inhibit amyloid assembly (Feng et al., 2008; Young et al., 2015). BSA inhibits the activity of colloidal small-molecule aggregates via adsorption (McGovern et al., 2002; Coan and Shoichet, 2007; Feng et al., 2008). Additionally, colloidal small-molecule aggregates can be precleared via centrifugation (McGovern et al., 2003). Thus, we used these two techniques to test whether inhibition of PAP248-286, PAP85-120, or SEM1(45-107) aggregation could be mediated via unexpected colloid formation by CLR01. CLR01 was found to inhibit PAP248-286, PAP85-120, and SEM1(45-107) fibril assembly in the presence of BSA (10 mg/ml) or when solutions containing CLR01 were centrifuged at 16,100×g for 20 min before adding the supernatant solution to the proteins (Figure 2—figure supplement 4). Thus, CLR01 is not inhibiting PAP248-286, PAP85-120, and SEM1(45-107) fibril assembly via a mechanism that involves colloidal CLR01 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)

BSA or preclearing CLR01 has no effect on the ability of CLR01 to inhibit formation of seminal amyloid fibrils.PAP248-286 (1 mM), PAP85-120 (1 mM), or SEM1(45-107) (500 µM) was incubated with CLR01 (1 mM) or buffer in the presence or absence of BSA (10 mg/ml) and agitated at 1400 rpm at 37°C for 68 hr. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibrillization was assessed via ThT fluorescence. 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 alone control to the other conditions (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.008
© Copyright Policy
Related In: Results  -  Collection

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

fig2s4: BSA or preclearing CLR01 has no effect on the ability of CLR01 to inhibit formation of seminal amyloid fibrils.PAP248-286 (1 mM), PAP85-120 (1 mM), or SEM1(45-107) (500 µM) was incubated with CLR01 (1 mM) or buffer in the presence or absence of BSA (10 mg/ml) and agitated at 1400 rpm at 37°C for 68 hr. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibrillization was assessed via ThT fluorescence. 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 alone control to the other conditions (*** denotes p < 0.0001).DOI:http://dx.doi.org/10.7554/eLife.05397.008
Mentions: Some small molecules must form higher order colloidal aggregates to inhibit amyloid assembly (Feng et al., 2008; Young et al., 2015). BSA inhibits the activity of colloidal small-molecule aggregates via adsorption (McGovern et al., 2002; Coan and Shoichet, 2007; Feng et al., 2008). Additionally, colloidal small-molecule aggregates can be precleared via centrifugation (McGovern et al., 2003). Thus, we used these two techniques to test whether inhibition of PAP248-286, PAP85-120, or SEM1(45-107) aggregation could be mediated via unexpected colloid formation by CLR01. CLR01 was found to inhibit PAP248-286, PAP85-120, and SEM1(45-107) fibril assembly in the presence of BSA (10 mg/ml) or when solutions containing CLR01 were centrifuged at 16,100×g for 20 min before adding the supernatant solution to the proteins (Figure 2—figure supplement 4). Thus, CLR01 is not inhibiting PAP248-286, PAP85-120, and SEM1(45-107) fibril assembly via a mechanism that involves colloidal CLR01 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