Förstner Philip, Bayer Fabienne, Kalu Nnanya, Felsen Susanne, Förtsch Christina, Aloufi Abrar, Ng David Y W, Weil Tanja, Nestorovich Ekaterina M, Barth Holger
Institute of Pharmacology and Toxicology, University of Ulm Medical Center , D-89081 Ulm, Germany.
Biomacromolecules. 2014 Jul 14;15(7):2461-74. doi: 10.1021/bm500328v. Epub 2014 Jul 2.
Dendrimers are unique highly branched macromolecules with numerous groundbreaking biomedical applications under development. Here we identified poly(amido amine) (PAMAM) dendrimers as novel blockers for the pore-forming B components of the binary anthrax toxin (PA63) and Clostridium botulinum C2 toxin (C2IIa). These pores are essential for delivery of the enzymatic A components of the internalized toxins from endosomes into the cytosol of target cells. We demonstrate that at low μM concentrations cationic PAMAM dendrimers block PA63 and C2IIa to inhibit channel-mediated transport of the A components, thereby protecting HeLa and Vero cells from intoxication. By channel reconstitution and high-resolution current recording, we show that the PAMAM dendrimers obstruct transmembrane PA63 and C2IIa pores in planar lipid bilayers at nM concentrations. These findings suggest a new potential role for the PAMAM dendrimers as effective polyvalent channel-blocking inhibitors, which can protect human target cells from intoxication with binary toxins from pathogenic bacteria.
树枝状大分子是独特的高度分支的大分子,目前有许多开创性的生物医学应用正在研发中。在这里,我们确定聚酰胺胺(PAMAM)树枝状大分子是二元炭疽毒素(PA63)和肉毒梭菌C2毒素(C2IIa)的成孔B组分的新型阻滞剂。这些孔对于将内化毒素的酶促A组分从内体递送到靶细胞的细胞质中至关重要。我们证明,在低 microM 浓度下,阳离子PAMAM树枝状大分子可阻断PA63和C2IIa,以抑制A组分的通道介导转运,从而保护HeLa和Vero细胞免受中毒。通过通道重建和高分辨率电流记录,我们表明PAMAM树枝状大分子在nM浓度下可阻断平面脂质双分子层中的跨膜PA63和C2IIa孔。这些发现表明PAMAM树枝状大分子作为有效的多价通道阻断抑制剂具有新的潜在作用,可保护人类靶细胞免受病原菌二元毒素的中毒。