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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 dose-dependently antagonize CLR01 binding to SEVI.SEVI fibrils were mixed with CLR01 and increasing amounts of lysine (1, 10 and 100-fold excess over CLR01) or poly-L-lysine (0.02, 0.25 and 2.5-fold excess of lysine monomer equivalents over CLR01) and centrifuged for 10 min at 20,000×g. The pellets were resuspended in 1 mM KCl and zeta potential was measured. Values represent means ±SD (n = 3).DOI:http://dx.doi.org/10.7554/eLife.05397.014
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fig4s1: Lysine and poly-L-lysine dose-dependently antagonize CLR01 binding to SEVI.SEVI fibrils were mixed with CLR01 and increasing amounts of lysine (1, 10 and 100-fold excess over CLR01) or poly-L-lysine (0.02, 0.25 and 2.5-fold excess of lysine monomer equivalents over CLR01) and centrifuged for 10 min at 20,000×g. The pellets were resuspended in 1 mM KCl and zeta potential was measured. Values represent means ±SD (n = 3).DOI:http://dx.doi.org/10.7554/eLife.05397.014

Mentions: Since the infectivity-enhancing activity of seminal amyloid fibrils is due to their positive surface charge (Roan et al., 2009; Arnold et al., 2012), we examined next whether CLR01 affected this property. To prevent fibril remodeling from occurring in these experiments, fibrils were mixed with CLR01, CLR03, or buffer, and samples were immediately centrifuged to remove unbound CLR01 and CLR03 molecules. Subsequently, the surface charge of resuspended fibrils was determined by zeta potential measurements. We found that CLR01, but not CLR03, neutralized the positive surface charge of SEVI, PAP85-120, and SEM1(49-107) fibrils within minutes (Figure 4A). Notably, pre-treatment of CLR01 with lysine or poly-L-lysine abrogated the fibril neutralizing activity of CLR01 (Figure 4—figure supplement 1), which further supports previous findings concerning the specificity underlying these interactions (Fokkens et al., 2005; Klärner et al., 2006, 2010). Next, confocal microscopy was used to assess whether this neutralization could abrogate fibril binding to YFP-tagged virions. As previously shown (Roan et al., 2011; Arnold et al., 2012; Yolamanova et al., 2013), buffer-treated fibrils efficiently sequestered virions (Figure 4B). In contrast, fibrils pretreated with CLR01, but not CLR03, were unable to form fibril–virion complexes (Figure 4B).10.7554/eLife.05397.013Figure 4.CLR01 neutralizes the positive surface charge of seminal amyloids and abrogates their ability to bind virions and enhance HIV infection.


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 dose-dependently antagonize CLR01 binding to SEVI.SEVI fibrils were mixed with CLR01 and increasing amounts of lysine (1, 10 and 100-fold excess over CLR01) or poly-L-lysine (0.02, 0.25 and 2.5-fold excess of lysine monomer equivalents over CLR01) and centrifuged for 10 min at 20,000×g. The pellets were resuspended in 1 mM KCl and zeta potential was measured. Values represent means ±SD (n = 3).DOI:http://dx.doi.org/10.7554/eLife.05397.014
© Copyright Policy
Related In: Results  -  Collection

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

fig4s1: Lysine and poly-L-lysine dose-dependently antagonize CLR01 binding to SEVI.SEVI fibrils were mixed with CLR01 and increasing amounts of lysine (1, 10 and 100-fold excess over CLR01) or poly-L-lysine (0.02, 0.25 and 2.5-fold excess of lysine monomer equivalents over CLR01) and centrifuged for 10 min at 20,000×g. The pellets were resuspended in 1 mM KCl and zeta potential was measured. Values represent means ±SD (n = 3).DOI:http://dx.doi.org/10.7554/eLife.05397.014
Mentions: Since the infectivity-enhancing activity of seminal amyloid fibrils is due to their positive surface charge (Roan et al., 2009; Arnold et al., 2012), we examined next whether CLR01 affected this property. To prevent fibril remodeling from occurring in these experiments, fibrils were mixed with CLR01, CLR03, or buffer, and samples were immediately centrifuged to remove unbound CLR01 and CLR03 molecules. Subsequently, the surface charge of resuspended fibrils was determined by zeta potential measurements. We found that CLR01, but not CLR03, neutralized the positive surface charge of SEVI, PAP85-120, and SEM1(49-107) fibrils within minutes (Figure 4A). Notably, pre-treatment of CLR01 with lysine or poly-L-lysine abrogated the fibril neutralizing activity of CLR01 (Figure 4—figure supplement 1), which further supports previous findings concerning the specificity underlying these interactions (Fokkens et al., 2005; Klärner et al., 2006, 2010). Next, confocal microscopy was used to assess whether this neutralization could abrogate fibril binding to YFP-tagged virions. As previously shown (Roan et al., 2011; Arnold et al., 2012; Yolamanova et al., 2013), buffer-treated fibrils efficiently sequestered virions (Figure 4B). In contrast, fibrils pretreated with CLR01, but not CLR03, were unable to form fibril–virion complexes (Figure 4B).10.7554/eLife.05397.013Figure 4.CLR01 neutralizes the positive surface charge of seminal amyloids and abrogates their ability to bind virions and enhance HIV infection.

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