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Am J Physiol Lung Cell Mol Physiol (November 1, 2002). doi:10.1152/ajplung.00316.2002
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Articles in PresS, published online ahead of print November 1, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00316.2002
Submitted on September 17, 2002
Accepted on October 30, 2002

Subcellular Localization of Pseudomonas Pyocyanin Cytotoxicity in Human Lung Epithelial Cells

Yunxia Q. O'Malley1, Maher Y. Abdalla2, Michael L. McCormick3, Krzysztof J. Reszka4, Gerene M. Denning1, and Bradley E. Britigan3*

1 Department of Internal Medicine and Research Service, VA Medical Center, Iowa City, Iowa, USA; Department of Internal Medicine, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA
2 Free Radical and Radiation Biology Program of the Department of Radiation Oncology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA
3 Department of Internal Medicine and Research Service, VA Medical Center, Iowa City, Iowa, USA; Department of Internal Medicine, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA; Free Radical and Radiation Biology Program of the Department of Radiation Oncology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA
4 Department of Internal Medicine and Research Service, VA Medical Center, Iowa City, Iowa, USA; Free Radical and Radiation Biology Program of the Department of Radiation Oncology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA

* To whom correspondence should be addressed. E-mail: bradley-britigan{at}uiowa.edu.

The Pseudomonas aeruginosa secretory product pyocyanin damages lung epithelium, likely due to redox cycling of pyocyanin and resultant superoxide and H2O2 generation. Subcellular site(s) of pyocyanin redox cycling and toxicity have not been well studied. Therefore, pyocyanin's effects on subcellular parameters in the A549 human type II alveolar epithelial cell line were examined. Confocal and electron microscopy studies suggested mitochondrial redox cycling of pyocyanin and extracellular H2O2 release, respectively. Pyocyanin decreased mitochondrial and cytoplasmic aconitase activity, ATP levels, cellular reduction of MTT, and mitochondrial membrane potential. These effects were transient at low pyocyanin concentrations and were linked to apparent cell-mediated metabolism of pyocyanin. Overexpression of MnSOD, but not CuZnSOD or catalase, protected cellular aconitase, but not ATP, from pyocyanin-mediated depletion. This suggests that loss of aconitase activity is not responsible for ATP depletion. How pyocyanin leads to ATP depletion, the mechanism of cellular metabolism of pyocyanin, and the impact of mitochondrial pyocyanin redox cycling on other cellular events are important areas for future study.




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