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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 remodel SEVI and PAP85-120 fibrils.SEVI (left) or PAP85-120 fibrils (right, 20 µM monomer) were treated with buffer or CLR01 (100 µM) in the presence or absence of BSA (10 mg/ml) for 2 hr at 37°C. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibril remodeling was then assessed by 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.05397.012
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fig3s2: BSA or preclearing CLR01 has no effect on the ability of CLR01 to remodel SEVI and PAP85-120 fibrils.SEVI (left) or PAP85-120 fibrils (right, 20 µM monomer) were treated with buffer or CLR01 (100 µM) in the presence or absence of BSA (10 mg/ml) for 2 hr at 37°C. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibril remodeling was then assessed by 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.05397.012

Mentions: To elucidate the role of lysine- and arginine-tweezer interactions in CLR01-mediated remodeling of SEVI and PAP85-120 fibrils, experiments were performed with fibrils comprised of PAP248-286(Ala) or PAP85-120(Ala). CLR01 was unable to remodel preformed SEVI(Ala) or PAP85-120(Ala) fibrils (Figure 3I–L). Indeed, we have been unable to establish conditions (e.g., higher CLR01 concentrations) where CLR01 remodels SEVI(Ala) or PAP85-120(Ala) fibrils (data not shown). Furthermore, addition of excess free lysine or poly-L-lysine to CLR01 prevented its SEVI and PAP85-120 amyloid-remodeling activity (Figure 3—figure supplement 1). Thus, CLR01-lysine, CLR01-arginine, or both contacts are critical for the remodeling process. By contrast, CLR01 retained SEVI and PAP85-120 amyloid-remodeling activity in the presence of BSA (10 mg/ml) or if CLR01 solutions were precleared via centrifugation (Figure 3—figure supplement 2). Thus, colloidal CLR01 aggregates do not contribute to the amyloid-remodeling activity of CLR01. Taken together, our findings suggest that the lysine- and arginine-specific molecular tweezer CLR01 prevents formation of seminal amyloid and remodels mature PAP85-120 and SEVI fibrils via binding to lysine and arginine residues.


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 remodel SEVI and PAP85-120 fibrils.SEVI (left) or PAP85-120 fibrils (right, 20 µM monomer) were treated with buffer or CLR01 (100 µM) in the presence or absence of BSA (10 mg/ml) for 2 hr at 37°C. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibril remodeling was then assessed by 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.05397.012
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Related In: Results  -  Collection

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fig3s2: BSA or preclearing CLR01 has no effect on the ability of CLR01 to remodel SEVI and PAP85-120 fibrils.SEVI (left) or PAP85-120 fibrils (right, 20 µM monomer) were treated with buffer or CLR01 (100 µM) in the presence or absence of BSA (10 mg/ml) for 2 hr at 37°C. In some reactions, CLR01 was first cleared by centrifugation at 16,100×g for 20 min at 25°C (precleared). Fibril remodeling was then assessed by 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.05397.012
Mentions: To elucidate the role of lysine- and arginine-tweezer interactions in CLR01-mediated remodeling of SEVI and PAP85-120 fibrils, experiments were performed with fibrils comprised of PAP248-286(Ala) or PAP85-120(Ala). CLR01 was unable to remodel preformed SEVI(Ala) or PAP85-120(Ala) fibrils (Figure 3I–L). Indeed, we have been unable to establish conditions (e.g., higher CLR01 concentrations) where CLR01 remodels SEVI(Ala) or PAP85-120(Ala) fibrils (data not shown). Furthermore, addition of excess free lysine or poly-L-lysine to CLR01 prevented its SEVI and PAP85-120 amyloid-remodeling activity (Figure 3—figure supplement 1). Thus, CLR01-lysine, CLR01-arginine, or both contacts are critical for the remodeling process. By contrast, CLR01 retained SEVI and PAP85-120 amyloid-remodeling activity in the presence of BSA (10 mg/ml) or if CLR01 solutions were precleared via centrifugation (Figure 3—figure supplement 2). Thus, colloidal CLR01 aggregates do not contribute to the amyloid-remodeling activity of CLR01. Taken together, our findings suggest that the lysine- and arginine-specific molecular tweezer CLR01 prevents formation of seminal amyloid and remodels mature PAP85-120 and SEVI fibrils via binding to lysine and arginine residues.

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