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Mechanism of Superoxide and Hydrogen Peroxide Formation by Fumarate Reductase, Succinate Dehydrogenase, and Aspartate Oxidase
Open accessAbstract
Oxidative stress is created in aerobic organisms when molecular oxygen chemically oxidizes redox enzymes, forming superoxide (O2*-) and hydrogen peroxide (H2O2). Prior work identified several flavoenzymes from Escherichia coli that tend to autoxidize. Of these, fumarate reductase (Frd) is notable both for its high turnover number and for its production of substantial O2*- in addition to H2O2. We have sought to identify characteristics of Frd that predispose it to this behavior. The ability of excess succinate to block autoxidation and the inhibitory effect of lowering the flavin potential indicate that all detectable autoxidation occurs from its FAD site, rather than from iron-sulfur clusters or bound quinones. The flavin adenine dinucleotide (FAD) moiety of Frd is unusually solvent-exposed, as evidenced by its ability to bind sulfite, and this may make it more likely to react adventitiously with O2*-. The autoxidizing species is apparently fully reduced flavin rather than flavosemiquinone, since treatments that more fully reduce the enzyme do not slow its turnover number. They do, however, switch the major product from O2*- to H2O2. A similar effect is achieved by lowering the potential of the proximal [2Fe-2S] cluster. These data suggest that Frd releases O2*- into bulk solution if this cluster is available to sequester the semiquinone electron; otherwise, that electron is rapidly transferred to the nascent superoxide, and H2O2 is the product that leaves the active site. This model is supported by the behavior of "aspartate oxidase" (aspartate:fumarate oxidoreductase), an Frd homologue that lacks Fe-S clusters. Its dihydroflavin also reacts avidly with oxygen, and H2O2 is the predominant product. In contrast, succinate dehydrogenase, with high potential clusters, generates O2*- exclusively. The identities of enzyme autoxidation products are significant because O2*- and H2O2 damage cells in different ways.
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In the citing paper
“Both enzymes produced ROS with a linear dependence on the dissolved oxygen concentration, although SQR produced ROS at a much lower rate.”
From Differential effects of complex II on mitochondrial ROS production and their relation to cardioprotective pre- and postconditioning (2013) · cites this paper as [101]
In the citing paper
“E. coli SQR exclusively produces superoxide.In contrast, E. coli QFR produces mainly superoxide at lower succinate concentrations and comparably more hydrogen peroxide at higher substrate concentrations [101].”
From Differential effects of complex II on mitochondrial ROS production and their relation to cardioprotective pre- and postconditioning (2013) · cites this paper as [101]
In the citing paper
“While a contribution of the Q site might occur in the respective mutants, the most likely source of ROS in wild-type complex II is the fully reduced FAD [37,101].”
From Differential effects of complex II on mitochondrial ROS production and their relation to cardioprotective pre- and postconditioning (2013) · cites this paper as [101]
In the citing paper
“Messner and Imlay concluded that QFR releases superoxide from the fully reduced flavin if the [2Fe-2S] cluster is available to sequester the semiquinone electron; otherwise this electron is rapidly transferred to the nascent superoxide and hydrogen peroxide is released [101].”
From Differential effects of complex II on mitochondrial ROS production and their relation to cardioprotective pre- and postconditioning (2013) · cites this paper as [101]
In the citing paper
“In principle, fully reduced FAD can either transfer one electron onto molecular oxygen which leads to the formation of superoxide (probably the major reaction in complex II, but only confirmed for E. coli SQR [101]) or it may also transfer both electrons onto O 2 which leads to the formation of H 2 O 2 (as shown for E. coli QFR [101]) (Fig. 2B).”
From Differential effects of complex II on mitochondrial ROS production and their relation to cardioprotective pre- and postconditioning (2013) · cites this paper as [101]
In the citing paper
“An impairment in the normal removal of electrons from the flavin group in SdhA could promote superoxide generation through the autoxidation of the reduced flavin group by O 2 in the matrix (34, 38, 46, 51).”
From Loss of the SdhB, but Not the SdhA, Subunit of Complex II Triggers Reactive Oxygen Species-Dependent Hypoxia-Inducible Factor Activation and Tumorigenesis (2008) · cites this paper as [34]
In the citing paper
“Some evidence suggests that the flavin adenine dinucleotide site in SdhA is the principal source of superoxide generation, due to the exposure of that moiety to the aqueous environment and the high electron density that develops when the electron flux downstream from that site is obstructed (34, 51).”
From Loss of the SdhB, but Not the SdhA, Subunit of Complex II Triggers Reactive Oxygen Species-Dependent Hypoxia-Inducible Factor Activation and Tumorigenesis (2008) · cites this paper as [34]
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Citations
Cites
Experiments in molecular genetics
Miller, J. H. (1972) · Medical Entomology and Zoology · Cited by 27,157
McCord, J. M., & Fridovich, I. (1969) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)63504-5 · Cited by 12,952
Zhou, M., Diwu, Z. J., Panchuk-Voloshina, N., & Haugland, R. P. (1997) · Analytical Biochemistry · doi:10.1006/abio.1997.2391 · Cited by 1,400
Storz, G., & Imlayt, J. A. (1999) · Current Opinion in Microbiology · doi:10.1016/s1369-5274(99)80033-2 · Cited by 1,077
Quantitative Aspects of the Production of Superoxide Anion Radical by Milk Xanthine Oxidase
Fridovich, I. (1970) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)62884-4 · Cited by 1,035
An Enzyme-Based Theory of Obligate Anaerobiosis: The Physiological Function of Superoxide Dismutase
McCord, J. M., Keele, B. B., & Fridovich, I. (1971) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.68.5.1024 · Cited by 973
Superoxide accelerates DNA damage by elevating free-iron levels
Keyer, K., & Imlay, J. A. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.24.13635 · Cited by 841
Self-citationActivation of molecular oxygen by flavins and flavoproteins.
Massey, V. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)31664-2 · Cited by 823
Seaver, L. C., & Imlay, J. A. (2001) · Journal of Bacteriology · doi:10.1128/jb.183.24.7173-7181.2001 · Cited by 797
Self-citationCarlioz, A., & Touati, D. (1986) · The EMBO Journal · doi:10.1002/j.1460-2075.1986.tb04256.x · Cited by 784
A mutation in succinate dehydrogenase cytochrome b causes oxidative stress and ageing in nematodes.
Ishii, N., Fujii, M., Hartman, P. S., Tsuda, M., Yasuda, K., Senoo-Matsuda, N., Yanase, S., Ayusawa, D., & Suzuki, K. (1998) · Nature · doi:10.1038/29331 · Cited by 687
Role of ubiquinone in the mitochondrial generation of hydrogen peroxide
Boveris, A., Cadenas, E., & Stoppani, A. O. M. (1976) · Biochemical Journal · doi:10.1042/bj1560435 · Cited by 655
The inactivation of Fe-S cluster containing hydro-lyases by superoxide
Flint, D. H., Tuminello, J. F., & Emptage, M. H. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)41538-4 · Cited by 637
Superoxide sensitivity of the Escherichia coli aconitase.
Gardner, P. R., & Fridovich, I. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)55001-8 · Cited by 514
Massey, V., & Palmer, G. (1966) · Biochemistry · doi:10.1021/bi00874a016 · Cited by 452
Massey, V., Strickland, S., Mayhew, S. G., Howell, L. G., Engel, P. C., Matthews, R. G., Schuman, M., & Sullivan, P. A. (1969) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(69)90287-3 · Cited by 426
The role of O2.− in the production of HO.: in vitro and in vivo
Liochev, S. I., & Fridovich, I. (1994) · Free Radical Biology and Medicine · doi:10.1016/0891-5849(94)90239-9 · Cited by 423
Assay of metabolic superoxide production in Escherichia coli.
Imlay, J. A., & Fridovich, I. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)89596-9 · Cited by 420
Self-citationStructure of the Escherichia coli Fumarate Reductase Respiratory Complex
Iverson, T. M., Luna-Chavez, C., Cecchini, G., & Rees, D. C. (1999) · Science · doi:10.1126/science.284.5422.1961 · Cited by 416
The Reactivity of Flavoproteins with Sulfite
Massey, V., Müller, F., Feldberg, R. S., Schuman, M., Sullivan, P. A., Howell, L. G., Mayhew, S. G., Matthews, R. G., & Foust, G. P. (1969) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)36376-7 · Cited by 343
Messner, K., & Imlay, J. A. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.15.10119 · Cited by 300
Self-citationIron–sulfur proteins with nonredox functions
Flint, D. H., & Allen, R. M. (1996) · Chemical Reviews · doi:10.1021/cr950041r · Cited by 280
Succinate dehydrogenase and fumarate reductase from Escherichia coli
Cecchini, G., Schröder, I., Gunsalus, R. P., & Maklashina, E. (2002) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/s0005-2728(01)00238-9 · Cited by 279
Senoo-Matsuda, N., Yasuda, K., Tsuda, M., Ohkubo, T., Yoshimura, S., Nakazawa, H., Hartman, P. S., & Ishii, N. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m104718200 · Cited by 271
Superoxide sensitivity of the Escherichia coli 6-phosphogluconate dehydratase
Gardner, P. R., & Fridovich, I. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)52319-x · Cited by 264
Superoxide and the production of oxidative DNA damage
Keyer, K., Gort, A. S., & Imlay, J. A. (1995) · Journal of Bacteriology · doi:10.1128/jb.177.23.6782-6790.1995 · Cited by 249
Self-citationalpha, beta-Dihydroxyisovalerate dehydratase. A superoxide-sensitive enzyme.
Kuo, C. F., Mashino, T., & Fridovich, I. (1987) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)61255-4 · Cited by 247
Fumarase C, the stable fumarase of Escherichia coli, is controlled by the soxRS regulon.
Liochev, S. I., & Fridovich, I. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.13.5892 · Cited by 237
REACTIVE OXYGEN INTERMEDIATES IN BIOCHEMISTRY
Naqui, A., Chance, B., & Cadenas, E. (1986) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.55.070186.001033 · Cited by 234
Studies on the oxidative half-reaction of xanthine oxidase.
Hille, R., & Massey, V. (1981) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)52512-1 · Cited by 160
Maklashina, E., Berthold, D. A., & Cecchini, G. (1998) · Journal of Bacteriology · doi:10.1128/jb.180.22.5989-5996.1998 · Cited by 141
The reaction of reduced xanthine oxidase with oxygen. Kinetics of peroxide and superoxide formation.
Porras, A. G., Olson, J. S., & Palmer, G. (1981) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)52513-3 · Cited by 139
Visualization of catalase on acrylamide gels
Gregory, E. M., & Fridovich, I. (1974) · Analytical Biochemistry · doi:10.1016/0003-2697(74)90440-0 · Cited by 134
A Metabolic Enzyme That Rapidly Produces Superoxide, Fumarate Reductase of Escherichia coli
Imlay, J. A. (1995) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)94548-5 · Cited by 132
Self-citationSuperoxide dismutase: “Positive” spectrophotometric assays
Misra, H. P., & Fridovich, I. (1977) · Analytical Biochemistry · doi:10.1016/0003-2697(77)90429-8 · Cited by 122
Bruice, T. C. (1984) · Israel Journal of Chemistry · doi:10.1002/ijch.198400008 · Cited by 119
How does oxygen inhibit central metabolism in the obligate anaerobe Bacteroides thetaiotaomicron
Pan, N., & Imlay, J. A. (2001) · Molecular Microbiology · doi:10.1046/j.1365-2958.2001.02343.x · Cited by 113
Self-citationFumarate Reductase Mutants of Escherichia coli That Lack Covalently Bound Flavin
Blaut, M., Whittaker, K. M., Valdovinos, A., Ackrell, B. A. C., Gunsalus, R. P., & Cecchini, G. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)80039-4 · Cited by 107
Heering, H. A., Weiner, J. H., & Armstrong, F. A. (1997) · Journal of the American Chemical Society · doi:10.1021/ja9723242 · Cited by 105
Léger, C., Heffron, K., Pershad, H. R., Maklashina, E., Luna-Chavez, C., Cecchini, G., Ackrell, B. A. C., & Armstrong, F. A. (2001) · Biochemistry · doi:10.1021/bi010889b · Cited by 97
Guest, J. R. (1981) · Microbiology · doi:10.1099/00221287-122-2-171 · Cited by 93
Mattevi, A., Tedeschi, G., Bacchella, L., Coda, A., Negri, A. L., & Ronchi, S. (1999) · Structure · doi:10.1016/s0969-2126(99)80099-9 · Cited by 91
Flavin radicals: Chemistry and biochemistry
Müller, F. (1987) · Free Radical Biology and Medicine · doi:10.1016/0891-5849(87)90009-8 · Cited by 83
Structure of FAD-Bound l -Aspartate Oxidase: Insight into Substrate Specificity and Catalysis,
Bossi, R. T., Negri, A. L., Tedeschi, G., & Mattevi, A. (2002) · Biochemistry · doi:10.1021/bi015939r · Cited by 75
Werth, M. T., Cecchini, G., Manodori, A. B., Ackrell, B. A. C., Schröder, I., Gunsalus, R. P., & Johnson, M. K. (1990) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.87.22.8965 · Cited by 73
Condon, C., Cammack, R., Patil, D. S., & Owen, P. (1985) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)39384-5 · Cited by 70
Silvius, J. R., & McElhaney, R. N. (1980) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.77.3.1255 · Cited by 70
Kowal, A. T., Werth, M. T., Manodori, A. B., Cecchini, G., Schroeder, I., Gunsalus, R. P., & Johnson, M. K. (1995) · Biochemistry · doi:10.1021/bi00038a024 · Cited by 65
l ‐Aspartate Oxidase from Escherichia coli
Tedeschi, G., Negri, A. L., Mortarino, M., Ceciliani, F., Simonic, T., Faotto, L., & Ronchi, S. (1996) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1996.0427u.x · Cited by 57
L‐Aspartate Oxidase from Escherichia coli
Mortarino, M., Negri, A. L., Tedeschi, G., Simonic, T., Duga, S., Gassen, H. G., & Ronchi, S. (1996) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1996.0418u.x · Cited by 50
Expression of theE. coli nadBGene and Characterization of the Gene Product L-Aspartate Oxidase
SEIFERT, J., Norbert, K. U. N. Z., Flachmann, R., Läufer, A., Jany, K., & Gassen, H. G. (1990) · Biological Chemistry Hoppe-Seyler · doi:10.1515/bchm3.1990.371.1.239 · Cited by 44
Miki, T., Yoshida, L. S., & Kakinuma, K. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)37017-6 · Cited by 43
Interactions of oxaloacetate with Escherichia coli fumarate reductase
Ackrell, B. A. C., Cochran, B. C., & Cecchini, G. (1989) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(89)90561-4 · Cited by 39
Singer, T. P., Salach, J. I., Hemmerich, P., & Ehrenberg, A. (1971) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(71)18099-8 · Cited by 37
[36] Fumarate reductase of Escherichia coli
Lemire, B. D., & Weiner, J. H. (1986) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(86)26038-3 · Cited by 32
Analyzing your complexes: structure of the quinol-fumarate reductase respiratory complex
Iverson, T. M., Luna-Chavez, C., Schröder, I., Cecchini, G., & Rees, D. C. (2000) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(00)00113-5 · Cited by 28
Activation of Succinate Dehydrogenase by FMNH2 and by Photoreduction
Salach, J. I., & Singer, T. P. (1974) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)42539-8 · Cited by 9
Research works printed but not linked: 2
- Ackrell, 1992, 3, 229
- Guest, 1981, J. Gen. Microbiol., 122, 171
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BackgroundImlay, J. A. (2003) · Annual Review of Microbiology · doi:10.1146/annurev.micro.57.030502.090938 · Cited by 2,201
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McEvoy, J. P., & Brudvig, G. W. (2006) · Chemical Reviews · doi:10.1021/cr0204294 · Cited by 1,542
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Brand, M. D. (2010) · Experimental Gerontology · doi:10.1016/j.exger.2010.01.003 · Cited by 1,217
“The rates during succinate oxidation by fumarate reductase, which catalyses the reverse reaction in Escherichia coli and lacks this mechanism, are much greater (Messner and Imlay, 2002).”
Background
Metadata
- Authors
- Messner, K. R., & Imlay, J. A.
- Affiliations
- University of Illinois Urbana-Champaign (US)
- Corresponding author
- Imlay, James A.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2002-11
- Volume
- 277
- Issue
- 45
- Pages
- 42563-42571
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2044412059
- Cited by
- 301
- References
- 57
- Citations by year
- 2026: 9 · 2025: 4 · 2024: 12 · 2023: 11 · 2022: 15 · 2021: 14 · 2020: 18 · 2019: 13 · 2018: 17 · 2017: 14 · 2016: 8 · 2015: 12 · 2014: 16 · 2013: 21 · 2012: 18
- FWCI
- 3.36
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (GM49640)
- National Institute of General Medical Sciences (R01GM049640, GM 49640, T32 GM07283-20, T32GM007283)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Metalloenzymes and iron-sulfur proteins
- Keywords
- fumarate reductase, succinate dehydrogenase, superoxide production, hydrogen peroxide production, FAD, flavin semiquinone, [2Fe-2S] cluster, oxidative stress, Escherichia coli
- MeSH terms
- Amino Acid Oxidoreductases, Cell Line, Cell Membrane, Escherichia coli, Flavin-Adenine Dinucleotide, Hydrogen Peroxide, Iron-Sulfur Proteins, Kinetics, Oxidation-Reduction, Solvents, Succinate Dehydrogenase, Superoxides, D-Aspartate Oxidase
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