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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

Lysine and poly-L-lysine antagonize the ability of CLR01 to inhibit spontaneous formation of seminal amyloid fibrils.(A) PAP248-286 (1 mM) was incubated with buffer or CLR01 (100 µM) in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (B) PAP248-286 (100 µM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (C) SEM1(45-107) (0.5 mM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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.05397.007
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fig2s3: Lysine and poly-L-lysine antagonize the ability of CLR01 to inhibit spontaneous formation of seminal amyloid fibrils.(A) PAP248-286 (1 mM) was incubated with buffer or CLR01 (100 µM) in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (B) PAP248-286 (100 µM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (C) SEM1(45-107) (0.5 mM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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.05397.007

Mentions: Next, we tested if an excess of free lysine or poly-L-lysine could interfere with the ability of CLR01 to block fibrillization of PAP248-286, PAP85-120, and SEM1(45-107) (Figure 2—figure supplement 3). Both 20 mM lysine and 1 mM poly-L-lysine completely abrogated CLR01 inhibition of fibril formation by these three peptides. These results strongly support direct interaction of the tweezer with lysine, arginine, or both residues in PAP248-286, PAP85-120, and SEM1(45-107) as the underlying mechanism of CLR01 inhibition. We suggest that these specific interactions preclude the conformational rearrangements necessary for amyloidogenesis.


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)

Lysine and poly-L-lysine antagonize the ability of CLR01 to inhibit spontaneous formation of seminal amyloid fibrils.(A) PAP248-286 (1 mM) was incubated with buffer or CLR01 (100 µM) in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (B) PAP248-286 (100 µM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (C) SEM1(45-107) (0.5 mM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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.05397.007
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fig2s3: Lysine and poly-L-lysine antagonize the ability of CLR01 to inhibit spontaneous formation of seminal amyloid fibrils.(A) PAP248-286 (1 mM) was incubated with buffer or CLR01 (100 µM) in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (B) PAP248-286 (100 µM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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). (C) SEM1(45-107) (0.5 mM) was incubated with CLR01 (100 µM) or buffer in the presence or absence of lysine (20 mM) or poly-L-lysine (1 mM) and agitated at 1400 rpm at 37°C for 68 hr. 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.05397.007
Mentions: Next, we tested if an excess of free lysine or poly-L-lysine could interfere with the ability of CLR01 to block fibrillization of PAP248-286, PAP85-120, and SEM1(45-107) (Figure 2—figure supplement 3). Both 20 mM lysine and 1 mM poly-L-lysine completely abrogated CLR01 inhibition of fibril formation by these three peptides. These results strongly support direct interaction of the tweezer with lysine, arginine, or both residues in PAP248-286, PAP85-120, and SEM1(45-107) as the underlying mechanism of CLR01 inhibition. We suggest that these specific interactions preclude the conformational rearrangements necessary for amyloidogenesis.

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