Superoxide Generation from Endothelial Nitric-oxide Synthase
Open accessAbstract
It has been previously shown that besides synthesizing nitric oxide (NO), neuronal and inducible NO synthase (NOS) generates superoxide (O-2) under conditions of L-arginine depletion. However, there is controversy regarding whether endothelial NOS (eNOS) can also produce O-2. Moreover, the mechanism and control of this process are not fully understood. Therefore, we performed electron paramagnetic resonance spin-trapping experiments to directly measure and characterize the O-2 generation from purified eNOS. With the spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), prominent signals of O-2 adduct, DMPO-OOH, were detected from eNOS in the absence of added tetrahydrobiopterin (BH4), and these were quenched by superoxide dismutase. This O-2 formation required Ca2+/calmodulin and was blocked by the specific NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME) but not its non-inhibitory enantiomer D-NAME. A parallel process of Ca2+/calmodulin-dependent NADPH oxidation was observed which was also inhibited by L-NAME but not D-NAME. Pretreatment of the enzyme with the heme blockers cyanide or imidazole also prevented O-2 generation. BH4 exerted dose-dependent inhibition of the O-2 signals generated by eNOS. Conversely, in the absence of BH4 L-arginine did not decrease this O-2 generation. Thus, eNOS can also catalyze O-2 formation, and this appears to occur primarily at the heme center of its oxygenase domain. O-2 synthesis from eNOS requires Ca2+/calmodulin and is primarily regulated by BH4 rather than L-arginine.
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In recent years, it has become clear that extracellular l-arginine and ADMA concentrations, compartmentalization, and membrane transporters are important for NOS function.
“However, in recent years, it has become clear that extracellular l-arginine and ADMA concentrations, compartmentalization, and membrane transporters are important for NOS function, as will be discussed below in sect. IIIB4 (167, 668, 1030).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [1030]
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Cites
Bradford, M. M. (1976) · Analytical Biochemistry · doi:10.1016/0003-2697(76)90527-3 · Cited by 255,302 · see citing works
Nitric oxide: physiology, pathophysiology, and pharmacology.
Moncada, S., Palmer, R. M., & Higgs, E. A. (1991) · Pharmacological Reviews · doi:10.1016/s0031-6997(25)06663-3 · Cited by 15,525 · see citing works
Nitric oxide synthases: Roles, tolls, and controls
Nathan, C., & Xie, Q. W. (1994) · Cell · doi:10.1016/0092-8674(94)90266-6 · Cited by 2,920 · see citing works
Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase
Bredt, D. S., Hwang, P. M., Glatt, C. E., Lowenstein, C. J., Reed, R. R., & Snyder, S. H. (1991) · Nature · doi:10.1038/351714a0 · Cited by 2,354 · see citing works
Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum.
Bredt, D. S., & Snyder, S. H. (1989) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.86.22.9030 · Cited by 1,849 · see citing works
Nitric oxide synthase structure and mechanism
Marletta, M. A. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)31375-9 · Cited by 1,341 · see citing works
Nitric Oxide Synthases: Properties and Catalytic Mechanism
Griffith, O. W., & Stuehr, D. J. (1995) · Annual Review of Physiology · doi:10.1146/annurev.ph.57.030195.003423 · Cited by 1,311 · see citing works
Generation of superoxide by purified brain nitric oxide synthase
Pou, S., Pou, W. S., Bredt, D. S., Snyder, S. H., & Rosen, G. M. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)35745-4 · Cited by 750 · see citing works
Xia, Y., Dawson, V. L., Dawson, T. M., Snyder, S. H., & Zweier, J. L. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.13.6770 · Cited by 718 · see citing works
Self-citationSuperoxide and peroxynitrite generation from inducible nitric oxide synthase in macrophages
Xia, Y., & Zweier, J. L. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.13.6954 · Cited by 691 · see citing works
Self-citationSTRUCTURE-FUNCTION ASPECTS IN THE NITRIC OXIDE SYNTHASES
Stuehr, D. J. (1997) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev.pharmtox.37.1.339 · Cited by 547 · see citing works
Ca2+/calmodulin-dependent formation of hydrogen peroxide by brain nitric oxide synthase
Heinzel, B., John, M., Klatt, P., Böhme, E., & Mayer, B. (1992) · Biochemical Journal · doi:10.1042/bj2810627 · Cited by 530 · see citing works
Pritchard, K. A., Groszek, L., Smalley, D. M., Sessa, W. C., Wu, M., Villalon, P., Wolin, M. S., & Stemerman, M. B. (1995) · Circulation Research · doi:10.1161/01.res.77.3.510 · Cited by 471 · see citing works
Gross, S. S., & Levi, R. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)35667-9 · Cited by 443 · see citing works
Endothelial Nitric Oxide Synthase-Dependent Superoxide Generation from Adriamycin
Vasquez-Vivar, J. M., Pavel Martasek, E., Hogg, N., Masters, B. S. S., Pritchard, K. A., & Kalyanaraman, B. (1997) · Biochemistry · doi:10.1021/bi971475e · Cited by 366 · see citing works
Production of hydroxyl radical by decomposition of superoxide spin-trapped adducts.
Finkelstein, E., Rosen, G. M., & Rauckman, E. J. (1982) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)14604-x · Cited by 298 · see citing works
Tetrahydrobiopterin and Dysfunction of Endothelial Nitric Oxide Synthase in Coronary Arteries
Cosentino, F., & Katušić, Z. S. (1995) · Circulation · doi:10.1161/01.cir.91.1.139 · Cited by 294 · see citing works
Abu‐Soud, H. M., Yoho, L. L., & Stuehr, D. J. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)31597-7 · Cited by 249 · see citing works
Roubaud, V., Sankarapandi, S., Kuppusamy, P., Tordo, P., & Zweier, J. L. (1997) · Analytical Biochemistry · doi:10.1006/abio.1997.2067 · Cited by 186 · see citing works
Self-citationMasters, B. S. S., McMillan, K., Sheta, E. A., Nishimura, J. S., Roman, L. J., & Pavel Martasek, E. (1996) · The FASEB Journal · doi:10.1096/fasebj.10.5.8621055 · Cited by 180 · see citing works
LIST, B. M., KLÖSCH, B., VÖLKER, C., GORREN, A. C. F., SESSA, W. C., WERNER, E. R., KUKOVETZ, W. R., SCHMIDT, K., & MAYER, B. (1997) · Biochemical Journal · doi:10.1042/bj3230159 · Cited by 147 · see citing works
Direct measurement of nitric oxide generation from nitric oxide synthase
Xia, Y., & Zweier, J. L. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.23.12705 · Cited by 136 · see citing works
Self-citationAdenosine Deaminase Inhibition Prevents Free Radical-mediated Injury in the Postischemic Heart
Xia, Y., Khatchikian, G. S., & Zweier, J. L. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.17.10096 · Cited by 101 · see citing works
Self-citationChen, P. F., Tsai, A., & Wu, K. K. (1995) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1995.2579 · Cited by 86 · see citing works
Self-citationEndothelial Nitric-oxide Synthase
Chen, P.-F., Tsai, A.-L., Berka, V., & Wu, K. K. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.24.14631 · Cited by 84 · see citing works
Self-citation
Flagged first, then by citations
Research works printed but not linked: 4
- Xia, Y., Dawson, V. L., Dawson, T. M., Snyder, S. H., and Zweier, J. L. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 6770 -6774
- Xia, Y., and Zweier, J. L. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 6954 -6958
- Bredt, D. S., and Snyder, S. H. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 9030 -9033
- Xia, Y., and Zweier, J. L. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 12705-12710
Cited by
Sorted by citations
Landmesser, U., Dikalov, S., Price, S. R., McCann, L., Fukai, T., Holland, S. M., Mitch, W. E., & Harrison, D. G. (2003) · Journal of Clinical Investigation · doi:10.1172/jci200314172 · Cited by 1,660 · see citing works
Endothelial dysfunction: a multifaceted disorder (The Wiggers Award Lecture)
Félétou, M., & Vanhoutte, P. M. (2006) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00292.2006 · Cited by 818 · see citing works
Laursen, J. B., Somers, M., Kurz, S., McCann, L., Warnholtz, A., Freeman, B. A., Tarpey, M., Fukai, T., & Harrison, D. G. (2001) · Circulation · doi:10.1161/01.cir.103.9.1282 · Cited by 721 · see citing works
Interactions of Peroxynitrite, Tetrahydrobiopterin, Ascorbic Acid, and Thiols
Kuzkaya, N., Weissmann, N., Harrison, D. G., & Dikalov, S. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m302227200 · Cited by 692 · see citing works
Mollnau, H., Wendt, M., Szöcs, K., Lassègue, B., Schulz, E., Oelze, M., Li, H., Bodenschatz, M., August, M., Kleschyov, A. L., et al. (2002) · Circulation Research · doi:10.1161/01.res.0000012569.55432.02 · Cited by 676 · see citing works
Reactive Oxygen Species in the Regulation of Synaptic Plasticity and Memory
Massaad, C. A., & Klann, E. (2011) · Antioxidants & Redox Signaling · doi:10.1089/ars.2010.3208 · Cited by 635 · see citing works
S-glutathionylation uncouples eNOS and regulates its cellular and vascular function
Chen, C.-A., Wang, T.-Y., Varadharaj, S., Reyes, L. A., Hemann, C., Talukder, M. A. H., Chen, Y.-R., Druhan, L. J., & Zweier, J. L. (2010) · Nature · doi:10.1038/nature09599 · Cited by 594 · see citing works
Self-citationSources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation
Tejero, J., Shiva, S., & Gladwin, M. T. (2019) · Physiological Reviews · doi:10.1152/physrev.00036.2017 · Cited by 541 · see citing works
Zou, M.-H., Shi, C., & Cohen, R. A. (2002) · Journal of Clinical Investigation · doi:10.1172/jci0214442 · Cited by 494 · see citing works
Oxidative stress and diabetic cardiovascular complications
Jay, D., Hitomi, H., & Griendling, K. K. (2006) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2005.06.018 · Cited by 464 · see citing works
Fleming, I., Michaelis, U. R., Bredenkötter, D., Fisslthaler, B., Dehghani, F., Brandes, R. P., & Busse, R. (2001) · Circulation Research · doi:10.1161/01.res.88.1.44 · Cited by 440 · see citing works
Explaining the Phenomenon of Nitrate Tolerance
Münzel, T., Daiber, A., & Mülsch, A. (2005) · Circulation Research · doi:10.1161/01.res.0000184694.03262.6d · Cited by 424 · see citing works
Asymmetrical Dimethylarginine: The Über Marker?
Cooke, J. P. (2004) · Circulation · doi:10.1161/01.cir.0000126823.07732.d5 · Cited by 396 · see citing works
Endothelium-derived nitric oxide and vascular physiology and pathology
Arnal, J.-F., Dinh-Xuan, A.-T., Pueyo, M., Darblade, B., & Rami, J. (1999) · Cellular and Molecular Life Sciences · doi:10.1007/s000180050358 · Cited by 313 · see citing works
eNOS Uncoupling in Cardiovascular Diseases - the Role of Oxidative Stress and Inflammation
Karbach, S., Wenzel, P., Waisman, A., Munzel, T., & Daiber, A. (2014) · Current Pharmaceutical Design · doi:10.2174/13816128113196660748 · Cited by 307 · see citing works
Ozaki, M., Kawashima, S., Yamashita, T., Hirase, T., Namiki, M., Inoue, N., Hirata, K.-I., Yasui, H., Sakurai, H., Yoshida, Y., et al. (2002) · Journal of Clinical Investigation · doi:10.1172/jci0215215 · Cited by 268 · see citing works
Cooke, J. P. (2003) · Atherosclerosis Supplements · doi:10.1016/s1567-5688(03)00034-5 · Cited by 266 · see citing works
Rueckschloss, U., Quinn, M. T., Holtz, J., & Morawietz, H. (2002) · Arteriosclerosis, Thrombosis, and Vascular Biology · doi:10.1161/01.atv.0000035392.38687.65 · Cited by 216 · see citing works
Dumitrescu, C., Biondi, R., Xia, Y., Cardounel, A. J., Druhan, L. J., Ambrosio, G., & Zweier, J. L. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0702986104 · Cited by 192 · see citing works
Self-citationAnderson, H. D. I., Rahmutula, D., & Gardner, D. G. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m309552200 · Cited by 186 · see citing works
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Funk, S. D., Yurdagul, A., & Orr, A. W. (2012) · International Journal of Vascular Medicine · doi:10.1155/2012/569654 · Cited by 176 · see citing works
Cunnington, C., Van Assche, T., Shirodaria, C., Kylintireas, I., Lindsay, A. C., Lee, J. M., Antoniades, C., Margaritis, M., Lee, R., Cerrato, R., et al. (2012) · Circulation · doi:10.1161/circulationaha.111.038919 · Cited by 166 · see citing works
Nitric-oxide synthase: A cytochrome P450 family foster child
Gorren, A. C. F., & Mayer, B. (2007) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/j.bbagen.2006.08.019 · Cited by 129 · see citing works
The role of dopamine in inflammation-associated depression: mechanisms and therapeutic implications
Felger, J. C. (2016) · Current Topics in Behavioral Neurosciences · doi:10.1007/7854_2016_13 · Cited by 115 · see citing works
Effect of Vitamin C on the Availability of Tetrahydrobiopterin in Human Endothelial Cells
Baker, T. A., Milstien, S., & Katusic, Z. S. (2001) · Journal of Cardiovascular Pharmacology · doi:10.1097/00005344-200103000-00012 · Cited by 113 · see citing works
Ihlemann, N., Rask-Madsen, C., Perner, A., Dominguez, H., Hermann, T., Køber, L., & Torp-Pedersen, C. (2003) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00008.2003 · Cited by 113 · see citing works
Regulation of eNOS-Derived Superoxide by Endogenous Methylarginines
Druhan, L. J., Forbes, S. P., Pope, A. J., Chen, C.-A., Zweier, J. L., & Cardounel, A. J. (2008) · Biochemistry · doi:10.1021/bi702377a · Cited by 104 · see citing works
Self-citationVaradharaj, S., Kelly, O. J., Khayat, R. N., Kumar, P. S., Ahmed, N., & Zweier, J. L. (2017) · Frontiers in Cardiovascular Medicine · doi:10.3389/fcvm.2017.00064 · Cited by 101 · see citing works
Self-citationChen, C.-A., De Pascali, F., Basye, A., Hemann, C., & Zweier, J. L. (2013) · Biochemistry · doi:10.1021/bi400404s · Cited by 71 · see citing works
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Kuhlencordt, P. J., Hötten, S., Schödel, J., Rützel, S., Hu, K., Widder, J., Marx, A., Huang, P. L., & Ertl, G. (2006) · Arteriosclerosis, Thrombosis, and Vascular Biology · doi:10.1161/01.atv.0000223143.88128.19 · Cited by 67 · see citing works
Boslett, J., Hemann, C., Christofi, F. L., & Zweier, J. L. (2018) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00139.2017 · Cited by 65 · see citing works
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McGarrity, S., Halldórsson, H., Palsson, S., Johansson, P. I., & Rolfsson, Ó. (2016) · Frontiers in Cardiovascular Medicine · doi:10.3389/fcvm.2016.00010 · Cited by 12 · see citing works
- Authors
- Xia, Y., Tsai, A.-L., Berka, V., & Zweier, J. L.
- Affiliations
- Johns Hopkins University (US)
- Corresponding author
- Zweier, Jay L.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1998-10
- Volume
- 273
- Issue
- 40
- Pages
- 25804-25808
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2063024150
- Cited by
- 705
- References
- 25
- Citations by year
- 2026: 6 · 2025: 4 · 2024: 5 · 2023: 13 · 2022: 10 · 2021: 9 · 2020: 16 · 2019: 18 · 2018: 25 · 2017: 17 · 2016: 16 · 2015: 18 · 2014: 21 · 2013: 26 · 2012: 37
- FWCI
- 14.09
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL-38324, HL52315, P50HL052315)
- National Institute of General Medical Sciences (GM-44911, R01GM044911)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- endothelial nitric oxide synthase, superoxide production, tetrahydrobiopterin, L-arginine, calcium/calmodulin regulation, NADPH oxidation, L-NAME, superoxide dismutase, DMPO spin trapping, imidazole, cyanide
- MeSH terms
- Arginine, Biopterins, Calcium, Calmodulin, Cyclic N-Oxides, Electron Spin Resonance Spectroscopy, Enzyme Inhibitors, Free Radicals, Humans, NADP, Recombinant Proteins, Sodium Cyanide, Spin Labels, Stereoisomerism, Superoxides, Nitric Oxide Synthase, NG-Nitroarginine Methyl Ester
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