Limits...
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

Preclearing CLR01 via centrifugation does not affect antiviral activity.CLR01 (50 µM) was centrifuged at 20,000×g and 20°C for 20 min. HIV-1 92TH014 was incubated for 10 min with the supernatant (S) or the pellet fraction (P). As a control, virus was also incubated either with PBS (Buffer) or 50 µM CLR01 that was not centrifuged (NC). After infection of TZM-bl cells with these mixtures infection rates were measured 2 days post infection. Values represent mean β-galactosidase activities derived from triplicate infections ±SD (RLU/s: relative light units per second). Unpaired t-tests were used to compare the buffer control to each CLR01 condition at each poly-L-lysine concentration (ns denotes not significant; ** denotes p < 0.01; *** denotes p < 0.001).DOI:http://dx.doi.org/10.7554/eLife.05397.021
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4536748&req=5

fig5s4: Preclearing CLR01 via centrifugation does not affect antiviral activity.CLR01 (50 µM) was centrifuged at 20,000×g and 20°C for 20 min. HIV-1 92TH014 was incubated for 10 min with the supernatant (S) or the pellet fraction (P). As a control, virus was also incubated either with PBS (Buffer) or 50 µM CLR01 that was not centrifuged (NC). After infection of TZM-bl cells with these mixtures infection rates were measured 2 days post infection. Values represent mean β-galactosidase activities derived from triplicate infections ±SD (RLU/s: relative light units per second). Unpaired t-tests were used to compare the buffer control to each CLR01 condition at each poly-L-lysine concentration (ns denotes not significant; ** denotes p < 0.01; *** denotes p < 0.001).DOI:http://dx.doi.org/10.7554/eLife.05397.021

Mentions: Incubation of the tweezer in solutions containing up to 5% (sevenfold molar excess) coated or free BSA did not reduce the anti-HIV activity of CLR01 (Figure 5—figure supplements 2, 3). Similarly, the antiviral activity of CLR01 was not reduced when CLR01 solutions were precleared of any aggregates via centrifugation (Figure 5—figure supplement 4). Moreover, the pellet fraction did not possess antiviral activity (Figure 5—figure supplement 4). These data strongly suggest that colloidal CLR01 aggregates do not contribute to the direct antiviral activity of CLR01.


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)

Preclearing CLR01 via centrifugation does not affect antiviral activity.CLR01 (50 µM) was centrifuged at 20,000×g and 20°C for 20 min. HIV-1 92TH014 was incubated for 10 min with the supernatant (S) or the pellet fraction (P). As a control, virus was also incubated either with PBS (Buffer) or 50 µM CLR01 that was not centrifuged (NC). After infection of TZM-bl cells with these mixtures infection rates were measured 2 days post infection. Values represent mean β-galactosidase activities derived from triplicate infections ±SD (RLU/s: relative light units per second). Unpaired t-tests were used to compare the buffer control to each CLR01 condition at each poly-L-lysine concentration (ns denotes not significant; ** denotes p < 0.01; *** denotes p < 0.001).DOI:http://dx.doi.org/10.7554/eLife.05397.021
© Copyright Policy
Related In: Results  -  Collection

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

fig5s4: Preclearing CLR01 via centrifugation does not affect antiviral activity.CLR01 (50 µM) was centrifuged at 20,000×g and 20°C for 20 min. HIV-1 92TH014 was incubated for 10 min with the supernatant (S) or the pellet fraction (P). As a control, virus was also incubated either with PBS (Buffer) or 50 µM CLR01 that was not centrifuged (NC). After infection of TZM-bl cells with these mixtures infection rates were measured 2 days post infection. Values represent mean β-galactosidase activities derived from triplicate infections ±SD (RLU/s: relative light units per second). Unpaired t-tests were used to compare the buffer control to each CLR01 condition at each poly-L-lysine concentration (ns denotes not significant; ** denotes p < 0.01; *** denotes p < 0.001).DOI:http://dx.doi.org/10.7554/eLife.05397.021
Mentions: Incubation of the tweezer in solutions containing up to 5% (sevenfold molar excess) coated or free BSA did not reduce the anti-HIV activity of CLR01 (Figure 5—figure supplements 2, 3). Similarly, the antiviral activity of CLR01 was not reduced when CLR01 solutions were precleared of any aggregates via centrifugation (Figure 5—figure supplement 4). Moreover, the pellet fraction did not possess antiviral activity (Figure 5—figure supplement 4). These data strongly suggest that colloidal CLR01 aggregates do not contribute to the direct antiviral activity of CLR01.

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