Dissecting the role of ADAM10 as a mediator of Staphylococcus aureus α-toxin action
| Autor: | Nadja Hellmann, Paul Saftig, Rolf Postina, Amable J. Rivas, Matthias Husmann, Qianqian Qin, Christian Hamm, Stefan Klein, Gisela von Hoven, Sabine Füser, Claudia Neukirch, Martina Meyenburg |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Staphylococcus aureus ADAM10 Bacterial Toxins medicine.disease_cause Biochemistry Virulence factor ADAM10 Protein Hemolysin Proteins Mice 03 medical and health sciences Catalytic Domain medicine Disintegrin Animals Molecular Biology Furin Cells Cultured Mice Knockout Metalloproteinase biology Cadherin Cell Membrane Cell Biology Staphylococcal Infections Cadherins Cell biology 030104 developmental biology biology.protein Calcium Intracellular Protein Binding |
| Zdroj: | Biochemical Journal. 473:1929-1940 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bcj20160062 |
| Popis: | Staphylococcus aureus is a leading cause of bacterial infections in humans, including life-threatening diseases such as pneumonia and sepsis. Its small membrane-pore-forming α-toxin is considered an important virulence factor. By destroying cell–cell contacts through cleavage of cadherins, the metalloproteinase ADAM10 (a disintegrin and metalloproteinase 10) critically contributes to α-toxin-dependent pathology of experimental S. aureus infections in mice. Moreover, ADAM10 was proposed to be a receptor for α-toxin. However, it is unclear whether the catalytic activity or specific domains of ADAM10 are involved in mediating binding and/or subsequent cytotoxicity of α-toxin. Also, it is not known how α-toxin triggers ADAM10’s enzymatic activity, and whether ADAM10 is invariably required for all α-toxin action on cells. In the present study, we show that efficient cleavage of the ADAM10 substrate epithelial cadherin (E-cadherin) requires supra-cytotoxic concentrations of α-toxin, leading to significant increases in intracellular [Ca2+]; the fall in cellular ATP levels, typically following membrane perforation, became observable at far lower concentrations. Surprisingly, ADAM10 was dispensable for α-toxin-dependent xenophagic targeting of S. aureus, whereas a role for α-toxin attack on the plasma membrane was confirmed. The catalytic site of ADAM10, furin cleavage site, cysteine switch and intracellular domain of ADAM10 were not required for α-toxin binding and subsequent cytotoxicity. In contrast, an essential role for the disintegrin domain and the prodomain emerged. Thus, co-expression of the prodomain with prodomain-deficient ADAM10 reconstituted binding of α-toxin and susceptibility of ADAM10-deficient cells. The results of the present study may help to inform structural analyses of α-toxin–ADAM10 interactions and to design novel strategies to counteract S. aureus α-toxin action. |
| Databáze: | OpenAIRE |
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