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Dal Molin, Federica and Tonello , Fiorella and Ladant, Daniel and Zornetta, Irene and Zamparo, Ilaria and Di Benedetto , Giulietta and Zaccolo , Manuela and Montecucco , Cesare (2006) Cell entry and cAMP imaging of anthrax edema toxin. [Online journal papers]

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Abstract (english)

The entry and enzymatic activity of the anthrax edema factor (EF) in different cell types was studied by monitoring EF-induced changes in intracellular cAMP with biochemical and microscopic methods. cAMP was imaged in live cells, transfected with a fluorescence resonance energy transfer biosensor based on the protein kinase A regulatory and catalytic subunits fused to CFP and YFP, respectively. The cAMP biosensor was located either in the cytosol or was membrane-bound owing to the addition of a tag determining its myristoylation/palmitoylation. Real-time imaging of cells expressing the cAMP biosensors provided the time course of EF catalytic activity and an indication of its subcellular localization. Bafilomycin A1, an inhibitor of the vacuolar ATPase proton pump, completely prevented EF activity, even when added long after the toxin. The time course of ppearance of the adenylate cyclase activity and of bafilomycin A1 action suggests that EF enters the cytosol from late endosomes. EF remains associated to these compartments and its activity shows a perinuclear localization generating intracellular cAMP concentration gradients from the cell centre to the periphery.

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EPrint type:Online journal papers
Anno di Pubblicazione:02 November 2006
Key Words:anthrax, cAMP, cell entry, edema toxin, FRET imaging
Settori scientifico-disciplinari MIUR:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Struttura di riferimento:Dipartimenti > pre 2012 - Dipartimento di Scienze Biomediche Sperimentali
Codice ID:984
Depositato il:24 Feb 2008
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