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Assay of metabolic superoxide production in Escherichia coli.
Open accessAbstract
Superoxide production has been measured in subcellular fractions of SOD-deficient Escherichia coli provided with physiological reductants. Although cytosolic enzyme(s) do generate O2-., the larger portion is produced by autoxidation of components of the respiratory electron-transport chain. At 37 degrees C and with pO2, NADH, and NAD+ levels matching those in vivo, respiring membrane vesicles generate 3 O2-./10,000 electrons transferred. This corresponds to intracellular O2-. production, in glucose-fed cells, of 5 microM/s. The high SOD content of normal cells restricts O2-. accumulation to 2.10(-10) M, with a moderate gradient from the membrane to the center of the cell. SOD-deficient mutants achieve a much higher steady-state content of O2-.. Rates of superoxide-mediated inactivation of certain enzymes are sufficiently rapid that even 10(-10) M O2-. imposes a significant oxidative stress.
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Phillips, J. P., Campbell, S. D., Michaud, D., Charbonneau, M., & Hilliker, A. J. (1989) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.86.8.2761 · Cited by 389
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The Free Radical Theory of Aging Matures
BECKMAN, K. B., & AMES, B. N. (1998) · Physiological Reviews · doi:10.1152/physrev.1998.78.2.547 · Cited by 3,879
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Kohanski, M. A., Dwyer, D. J., Hayete, B., Lawrence, C. A., & Collins, J. J. (2007) · Cell · doi:10.1016/j.cell.2007.06.049 · Cited by 2,982
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Cadenas, E., & Davies, K. J. A. (2000) · Free Radical Biology and Medicine · doi:10.1016/s0891-5849(00)00317-8 · Cited by 3,010
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Imlay, J. A. (2008) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.77.061606.161055 · Cited by 1,563
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Metadata
- Authors
- Imlay, J. A., & Fridovich, I.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1991-04
- Volume
- 266
- Issue
- 11
- Pages
- 6957-6965
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1537686192
- Cited by
- 420
- References
- 1
- FWCI
- 4.69
- Type
- journal article
- Language
- en
- Open access
- hybrid
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