The aim of this study was to compare two methods for quantification of changes in intracellular potassium concentration (decrease from ∼140 to ∼20mM) due to the action of a pore-forming toxin, the adenylate cyclase toxin (CyaA) from the pathogenic bacterium Bordetella pertussis. CyaA was incubated with stably transfected K1 Chinese hamster ovary cells expressing the toxin receptor CD11b/CD18 and the decrease in potassium concentration in the cells was followed by inductively coupled plasma mass spectrometry (ICP-MS). It is shown that this method is superior in terms of sensitivity, accuracy, and temporal resolution over the method employing the potassium-binding benzofuran isophthalate-acetoxymethyl ester fluorescent indicator. The ICP-MS procedure was found to be a reliable and straightforward analytical approach enabling kinetic studies of CyaA action at physiologically relevant toxin concentrations (<1000ng/ml) in biological microsamples.

Institute of Analytical Chemistry Academy of Sciences of the Czech Republic 60200 Brno Czech Republic

Laboratory of Molecular Biology of Bacterial Pathogens Cell and Molecular Microbiology Division Institute of Microbiology Academy of Sciences of the Czech Republic 142 20 Prague 4 Czech Republic

Laboratory of Molecular Biology of Bacterial Pathogens Cell and Molecular Microbiology Division Institute of Microbiology Academy of Sciences of the Czech Republic 142 20 Prague 4 Czech Republic; Department of Genetics and Microbiology Faculty of Science Charles University Prague Prague 2 Czech Republic

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