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Quantification of potassium levels in cells treated with Bordetella adenylate cyclase toxin
T. Wald, I. Petry-Podgorska, R. Fiser, T. Matousek, J. Dedina, R. Osicka, P. Sebo, J. Masin,
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
ProQuest Central
from 2013-01-01 to 1 year ago
Medline Complete (EBSCOhost)
from 2003-01-01 to 1 year ago
Health & Medicine (ProQuest)
from 2013-01-01 to 1 year ago
- MeSH
- Adenylate Cyclase Toxin toxicity MeSH
- CD11b Antigen genetics MeSH
- CD18 Antigens genetics MeSH
- Bordetella pertussis enzymology MeSH
- CHO Cells MeSH
- Cricetulus MeSH
- Potassium chemistry metabolism MeSH
- Fluorescent Dyes chemistry MeSH
- Mass Spectrometry methods MeSH
- Intracellular Space drug effects metabolism MeSH
- Cricetinae MeSH
- Humans MeSH
- Transfection MeSH
- Animals MeSH
- Check Tag
- Cricetinae MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
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.
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- $a 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.
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