Cyclophilin-Facilitated Membrane Trans location as Pharmacological Target to Prevent Intoxication of Mammalian Cells by Binary Clostridial Actin ADP-Ribosylated Toxins

JOURNAL OF MOLECULAR BIOLOGY

卷:427

Article

Ernst, Katharina¹  Langer, Simon¹  Kaiser, Eva¹  Osseforth, Christian²  Michaelis, Jens²  Popoff, Michel R.³  Schwan, Carsten⁴  Aktories, Klaus⁴  Kahlert, Viktoria⁵  Malesevic, Miroslav⁶  Schiene-Fischer, Cordelia⁶  Barth, Holger¹ 

[1] Univ Ulm, Med Ctr, Inst Pharmacol & Toxicol, D-89081 Ulm, Germany

[2] Univ Ulm, Inst Biophys, D-89081 Ulm, Germany

[3] Inst Pasteur, Dept Anaerob Bacteria, F-75724 Paris, France

[4] Univ Freiburg, Inst Expt & Clin Pharmacol & Toxicol, D-79104 Freiburg, Germany

[5] Max Planck Res Unit Enzymol Prot Folding Halle, D-06120 Halle, Saale, Germany

[6] Univ Halle Wittenberg, Inst Biochem & Biotechnol, D-06120 Halle, Saale, Germany

关键词: bacterial protein toxin;    cellular uptake;    membrane transport;    cyclophilin 40;    PPlase;   

DOI  :  10.1016/j.jmb.2014.07.013

来源: Elsevier

【 摘 要 】

Clostridium botulinum C2 toxin, Clostridium perfringens iota toxin and Clostridium difficile CDT belong to the family of binary actin ADP-ribosylating toxins and are composed of a binding/translocation component and a separate enzyme component. The enzyme components ADP-ribosylate G-actin in the cytosol of target cells resulting in depolymerization of F-actin, cell rounding and cell death. The binding/translocation components bind to their cell receptors and form complexes with the respective enzyme components. After receptor-mediated endocytosis, the binding/translocation components form pores in membranes of acidified endosomes and the enzyme components translocate through these pores into the cytosol. This step is facilitated by the host cell chaperone heat shock protein 90 and peptidyl-prolyl cis/trans isomerases including cyclophilin A. Here, we demonstrate that a large isoform of cyclophilin A, the multi-domain enzyme cyclophilin 40 (Cyp40), binds to the enzyme components C2I, la and CDTa in vitro. Isothermal titration calorimetry revealed a direct binding to C2I with a calculated affinity of 101 nM and to la with an affinity of 1.01 mu M. Closer investigation for the prototypic C2I revealed that binding to Cyp40 did not depend on its ADP-ribosyltransferase activity but was stronger for unfolded C2I. The interaction of C2I with Cyp40 was also demonstrated in lysates from C2-treated cells by pull-down. Treatment of cells with a non-immunosuppressive cyclosporine A derivative, which still binds to and inhibits the peptidyl-prolyl cis/trans isomerase activity of cyclophilins, protected cells from intoxication with C2, iota and CDT toxins, offering an attractive approach for development of novel therapeutic strategies against binary actin ADP-ribosylating toxins. (C) 2014 Elsevier Ltd. All rights reserved.

【 授权许可】

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