Quantification of potassium levels in cells treated with Bordetella adenylate cyclase toxin

0428410 - MBÚ 2015 RIV US eng J - Článek v odborném periodiku
Wald, Tomáš - Petry-Podgórska, Inga - Fišer, Radovan - Matoušek, Tomáš - Dědina, Jiří - Osička, Radim - Šebo, Peter - Mašín, Jiří
Quantification of potassium levels in cells treated with Bordetella adenylate cyclase toxin.
Analytical Biochemistry. Roč. 450, APR 2014 (2014), s. 57-62. ISSN 0003-2697. E-ISSN 1096-0309
Grant CEP: GA ČR(CZ) GAP302/11/0580; GA ČR GA13-14547S; GA ČR GAP302/12/0460
Institucionální podpora: RVO:61388971 ; RVO:68081715
Klíčová slova: Potassium * Adenylate cyclase toxin * RTX
Kód oboru RIV: CE - Biochemie
Impakt faktor: 2.219, rok: 2014

The aim of this study was to compare two methods for quantification of changes in intracellular potassium concentration (decrease from similar to 140 to similar to 20 mM) 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 (<1000 ng/ml) in biological microsamples
Trvalý link: http://hdl.handle.net/11104/0233776