Nitric oxide, oxidants, and protein tyrosine nitration

Departamento de Bioquímica and Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, Avda. General Flores 2125, 11800 Montevideo, Uruguay

^E-mail:yu.ude.demf@idarr.

Communicated by Louis J. Ignarro, University of California School of Medicine, Los Angeles, CA, January 12, 2004

Received 2003 Nov 12

Abstract

The occurrence of protein tyrosine nitration under disease conditions is now firmly established and represents a shift from the signal transducing physiological actions of ^{NO to oxidative and potentially pathogenic pathways. Tyrosine nitration is mediated by reactive nitrogen species such as peroxynitrite anion (ONOO}^{) and nitrogen dioxide (}^NO₂), formed as secondary products of ^{NO metabolism in the presence of oxidants including superoxide radicals (} equation M1 ), hydrogen peroxide (H₂O₂), and transition metal centers. The precise interplay between ^{NO and oxidants and the identification of the proximal intermediate(s) responsible for nitration}in vivo have been under controversy. Despite the capacity of peroxynitrite to mediate tyrosine nitration in vitro, its role on nitration in vivo has been questioned, and alternative pathways, including the nitrite/H₂O₂/hemeperoxidase and transition metal-dependent mechanisms, have been proposed. A balanced analysis of existing evidence indicates that (i) different nitration pathways can contribute to tyrosine nitration in vivo, and (ii) most, if not all, nitration pathways involve free radical biochemistry with carbonate radicals ( equation M2 ) and/or oxo–metal complexes oxidizing tyrosine to tyrosyl radical followed by the diffusion-controlled reaction with ^NO₂ to yield 3-nitrotyrosine. Although protein tyrosine nitration is a low-yield process in vivo, 3-nitrotyrosine has been revealed as a relevant biomarker of ^{NO-dependent oxidative stress; additionally, site-specific nitration focused on particular protein tyrosines may result in modification of function and promote a biological effect. Tissue distribution and quantitation of protein 3-nitrotyrosine, recognition of the predominant nitration pathways and individual identification of nitrated proteins in disease states open new avenues for the understanding and treatment of human pathologies.}

Abstract

Early after the discovery of the signal transducing physiological functions of the free radical nitric oxide (^{NO) in the vasculature and nervous system (e.g., vasodilation and neurotransmission), it became evident that}^{NO could also participate as a cytotoxic effector molecule and/or a pathogenic mediator when produced at high rates by either inflammatory stimuli-induced nitric oxide synthase (iNOS) or overstimulation of the constitutive forms (eNOS and nNOS) (}1). Much of ^{NO-mediated pathogenicity depends on the formation of secondary intermediates such as peroxynitrite anion (ONOO}^{) and nitrogen dioxide (}^NO₂) that are typically more reactive and toxic than ^NOper se (2). The formation of reactive nitrogen species from ^{NO requires the presence of oxidants such as superoxide radicals (} equation M3 ), hydrogen peroxide (H₂O₂), and transition metal centers, the concentration of which can be increased either by ^{NO itself or by the same mediators that up-regulate}^{NO production. Nitrogen dioxide can also be formed in hydrophobic environments from the reactions of}^{NO with molecular oxygen, where these species concentrate (}3, 4). One of the molecular footprints left by the reactions of reactive nitrogen species with biomolecules is the nitration (i.e., addition of nitro group, –NO₂) of protein tyrosine residues to 3-nitrotyrosine. The formation of protein 3-nitrotyrosine was originally addressed in early protein chemistry studies with tetranitromethane aimed at establishing the function of tyrosines in proteins (5). This now-established posttranslational modification attracts considerable interest to biomedical research, because it can alter protein function, is associated to acute and chronic disease states, and can be a predictor of disease risk.

Seminal work by Beckman et al. (6) and Ischiropoulos et al. (7) demonstrated the capacity of peroxynitrite to cause protein tyrosine nitration in vitro and established the concept that biologically produced intermediates could promote nitration in vivo (8, 9), as was also suggested in an earlier work by Ohshima et al. (10). However, the role of peroxynitrite as a central species in biological nitration has been more recently questioned (11, 12), and alternative pathways, most notably mechanisms that depend on the formation of ^NO₂ by the action of hemeperoxidases and/or transition metal complexes on a main product of ^{NO metabolism, nitrite (} equation M4 ), have been presented as contributors (12–15).

In addition to nitration, inflammatory conditions promote other oxidative modifications in tyrosine such as chlorination, bromination, and hydroxylation to 3-chloro-, 3-bromo-, or 3-hydroxytyrosine, the detection of which may assist in identifying preferential nitration pathways. The extent of these oxidative modifications in vivo can reach similar values per milligram of tissue protein (e.g., 10–100 pmol/mg) and are comparable to the levels of protein S-nitrosation. It is also important to appreciate that other oxidative processes triggered by reactive nitrogen species such as thiol and methionine oxidation, disruption of iron–sulfur clusters, and oxidation of transition metal centers (2) can, in many cases, be more relevant than nitration in the promotion cell dysfunction/death.

A balanced analysis of all of the existing in vitro and in vivo evidence indicates that more than one pathway can contribute to protein tyrosine nitration. Interestingly, the alternative nitration pathways share common characteristics, because they involve free radical biochemistry with the participation of transient tyrosyl, ^NO₂, and carbonate ( equation M5 ) radicals and/or oxo–metal complexes.

Acknowledgments

I thank Drs. Stanley L. Hazen, Harry Ischiropoulos, Bruce A. Freeman, Joe S. Beckman, Irwin Fridovich, Jonathan S. Stamler, Jean Claude Drapier, and Ronald P. Mason for valuable comments, and my coinvestigators in Montevideo for contributions to the elucidation of various aspects presented in the manuscript. This work was supported by the Howard Hughes Medical Institute, Fogarty–National Institutes of Health, the John Simon Guggenheim Memorial Foundation, and Universidad de la República.

Acknowledgments

Notes

Abbreviations: NOS, NO synthase; MPO, myeloperoxidase; EPO, eosinophil peroxidase; SOD, superoxide dismutase; LMW, low molecular weight.

Notes

Abbreviations: NOS, NO synthase; MPO, myeloperoxidase; EPO, eosinophil peroxidase; SOD, superoxide dismutase; LMW, low molecular weight.

Footnotes

^{The debate on the biological production and measurement of peroxynitrite is analogous to a previous one on} equation M69 that arose after the discovery of SOD by J. M. McCord and I. Fridovich. The recurring discussions during the first decade led I. Fridovich and associates to invest large research efforts into fully (and redundantly) establishing the concept that equation M70 was a biologically relevant reactive intermediate.

^{Under some circumstances, SOD could augment the bioavailability of}^{NO by} equation M71 mechanisms such as (i) the SOD-mediated oxidation of nitroxyl anion (NO^{) to}^{NO (}26);. and (ii) the SOD-mediated reduction of S-nitrosothiols (27). These alternative mechanisms for the actions of SOD could mainly operate intracellularly (28). The immediate vascular responses to extracellularly added SOD are due to the prevention of the oxidative inactivation of readily diffusible ^NO. equation M72 interactions in the vasculature are viewed as contributions to the development of vascular disease states, including atherogenesis, hypertension, and hyperglycemia (29).

^{In the electrophilic aromatic nitration mechanism, there is no net redox change in the metal center (Eqs.}7–10). However, direct and unambiguous spectroscopic observation is sometimes difficult due to low steady-state concentrations and/or the absorption of peroxynitrite–metal complexes; additionally, there is no formation of ^NO₂.

^{Nitrated proteins can also undergo enhanced proteolytic degradation (}54), and reports have suggested that a denitrase activity may be present in vivo (55).

^{Some metabolic conditions favor the role of NOS as a significant source of peroxynitrite (}74). NOS, in turn, can be nitrated and potentially inactivated both in vitro and in vivo.

^{On the contrary, expression of amyotrophic lateral sclerosis-associated mutants of CuZnSOD leads to increased motoneuron nitration and apoptosis (}59), possibly due to a toxic gain of function of SOD that promotes peroxynitrite formation and nitration reactions.

^{Biological nitration is not restricted only to tyrosine nitration but may also result in nitration of tryptophan residues, as well as DNA bases, sugars, and lipids that can result in altered or new biological activities.}

Footnotes

References

1. Ignarro, L. J. (1990) Annu. Rev. Pharmacol. Toxicol.30, 535–560. [[PubMed]
2. Radi, R., Denicola, A., Alvarez, B., Ferrer-Sueta, G. & Rubbo, H. (2000) in Nitric Oxide, ed. Ignarro, L. (Academic, San Diego), pp. 57–82.
3. Liu, X., Miller, M. J., Joshi, M. S., Thomas, D. D. & Lancaster, J. R., Jr. (1998) Proc. Natl. Acad. Sci. USA95, 2175–2179.
4. Denicola, A., Batthyany, C., Lissi, E., Freeman, B. A., Rubbo, H. & Radi, R. (2002) J. Biol. Chem.277, 932–936. [[PubMed]
5. Sokolovsky, M., Riordan, J. F. & Vallee, B. L. (1966) Biochemistry5, 3582–3589. [[PubMed]
6. Beckman, J. S., Ischiropoulos, H., Zhu, L., van der Woerd, M., Smith, C., Chen, J., Harrison, J., Martin, J. C. & Tsai, M. (1992) Arch. Biochem. Biophys.298, 438–445. [[PubMed]
7. Ischiropoulos, H., Zhu, L., Chen, J., Tsai, M., Martin, J. C., Smith, C. D. & Beckman, J. S. (1992) Arch. Biochem. Biophys.298, 431–437. [[PubMed]
8. Ischiropoulos, H., Zhu, L. & Beckman, J. S. (1992) Arch. Biochem. Biophys.298, 446–451. [[PubMed]
9. Beckmann, J. S., Ye, Y. Z., Anderson, P. G., Chen, J., Accavitti, M. A., Tarpey, M. M. & White, C. R. (1994) Biol. Chem. Hoppe–Seyler375, 81–88. [[PubMed]
10. Ohshima, H., Friesen, M., Brouet, I. & Bartsch, H. (1990) Food Chem. Toxicol.28, 647–652. [[PubMed]
11. Pfeiffer, S., Schmidt, K. & Mayer, B. (2000) J. Biol. Chem.275, 6346–6352. [[PubMed]
12. Thomas, D. D., Espey, M. G., Vitek, M. P., Miranda, K. M. & Wink, D. A. (2002) Proc. Natl. Acad. Sci. USA99, 12691–12696.
13. Eiserich, J. P., Hristova, M., Cross, C. E., Jones, A. D., Freeman, B. A., Halliwell, B. & van der Vliet, A. (1998) Nature391, 393–397. [[PubMed]
14. Bian, K., Gao, Z., Weisbrodt, N. & Murad, F. (2003) Proc. Natl. Acad. Sci. USA100, 5712–5717.
15. Brennan, M. L., Wu, W., Fu, X., Shen, Z., Song, W., Frost, H., Vadseth, C., Narine, L., Lenkiewicz, E., Borchers, M. T., et al. (2002) J. Biol. Chem.277, 17415–17427. [[PubMed]
16. Fukuto, J. & Ignarro, L. (1997) Acc. Chem. Res.30, 149–152. [PubMed]
17. Eu, J. P., Liu, L., Zeng, M. & Stamler, J. S. (2000) Biochemistry39, 1040–1047. [[PubMed]
18. Denicola, A., Souza, J. M. & Radi, R. (1998) Proc. Natl. Acad. Sci. USA95, 3566–3571.
19. Radi, R., Peluffo, G., Alvarez, M. N., Naviliat, M. & Cayota, A. (2001) Free Radical Biol. Med.30, 463–488. [[PubMed]
20. Gryglewski, R. J., Palmer, R. M. & Moncada, S. (1986) Nature320, 454–456. [[PubMed]
21. Rubanyi, G. M. & Vanhoutte, P. M. (1986) Am. J. Physiol.250, H822–H827. [[PubMed]
22. Ignarro, L. J., Byrns, R. E., Buga, G. M., Wood, K. S. & Chaudhuri, G. (1988) J. Pharmacol. Exp. Ther.244, 181–189. [[PubMed]
23. Liochev, S. I. & Fridovich, I. (2002) Free Radical Biol. Med.33, 137–141. [[PubMed]
24. Buerk, D. G., Lamkin-Kennard, K. & Jaron, D. (2003) Free Radical Biol. Med.34, 1488–1503. [[PubMed]
25. Clark, S. R., Coffey, M. J., Maclean, R. M., Collins, P. W., Lewis, M. J., Cross, A. R. & O'Donnell, V. B. (2002) J. Immunol.169, 5889–5896. [[PubMed]
26. Murphy, M. E. & Sies, H. (1991) Proc. Natl. Acad. Sci. USA88, 10860–10864.
27. Jourd'heuil D., Laroux, F. S., Miles, A. M., Wink, D. A. & Grisham, M. B. (1999) Arch. Biochem. Biophys.361, 323–330. [[PubMed]
28. Hobbs, A. J., Fukuto, J. M. & Ignarro, L. (1994) Proc. Natl. Acad. Sci. USA91, 10992–10996.
29. Sowers, J. R. (2002) N. Engl. J. Med.346, 1999–2001. [[PubMed]
30. Gutierrez, H. H., Nieves, B., Chumley, P., Rivera, A. & Freeman, B. A. (1996) Free Radical Biol. Med.21, 43–52. [[PubMed]
31. Sarti, P., Avigliano, L., Gorlach, A. & Brune, B. (2002) Cell Death Differ.9, 1160–1162. [[PubMed]
32. Radi, R., Beckman, J. S., Bush, K. M. & Freeman, B. A. (1991) J. Biol. Chem.266, 4244–4250. [[PubMed]
33. Trujillo, M. & Radi, R. (2002) Arch. Biochem. Biophys.397, 91–98. [[PubMed]
34. Arteel, G. E., Briviba, K. & Sies, H. (1999) FEBS Lett.445, 226–230. [[PubMed]
35. Bryk, R., Griffin, P. & Nathan, C. (2000) Nature407, 211–215. [[PubMed]
36. Brito, C., Naviliat, M., Tiscornia, A. C., Vuillier, F., Gualco, G., Dighiero, G., Radi, R. & Cayota, A. M. (1999) J. Immunol.162, 3356–3366. [[PubMed]
37. Go, Y. M., Patel, R. P., Maland, M. C., Park, H., Beckman, J. S., Darley-Usmar, V. M. & Jo, H. (1999) Am. J. Physiol.277, H1647–H1653. [[PubMed]
38. Mihm, M. J., Wattanapitayakul, S. K., Piao, S. F., Hoyt, D. G. & Bauer, J. A. (2003) Biochem. Pharmacol.65, 1189–1197. [[PubMed]
39. Cosentino, F., Eto, M., De Paolis, P., van der Loo, B., Bachschmid, M., Ullrich, V., Kouroedov, A., Delli Gatti, C., Joch, H., Volpe, M., et al. (2003) Circulation107, 1017–1023. [[PubMed]
40. Augusto, O., Bonini, M. G., Amanso, A. M., Linares, E., Santos, C. C. & De Menezes, S. L. (2002) Free Radical Biol. Med.32, 841–859. [[PubMed]
41. Hazen, S. L., Gaut, J. P., Hsu, F. F., Crowley, J. R., d'Avignon, A. & Heinecke, J. W. (1997) J. Biol. Chem.272, 16990–16998. [[PubMed]
42. Fridovich, I(1997) J. Biol. Chem.272, 18515–18517. [[PubMed][Google Scholar]
43. Yamazaki, I. & Piette, L. H. (1990) J. Biol. Chem.265, 13589–13594. [[PubMed]
44. Burner, U., Furtmuller, P. G., Kettle, A. J., Koppenol, W. H. & Obinger, C. (2000) J. Biol. Chem.275, 20597–20601. [[PubMed]
45. Sampson, J. B., Rosen, H. & Beckman, J. S. (1996) Methods Enzymol.269, 210–218. [[PubMed]
46. Ferrer-Sueta, G., Quijano, C., Alvarez, B. & Radi, R. (2002) Methods Enzymol.349, 23–37. [[PubMed]
47. Bouton, C. & Drapier, J. C. (2003) Sci. STKE182, pe17. [[PubMed]
48. Soum, E., Brazzolotto, X., Goussias, C., Bouton, C., Moulis, J. M., Mattioli, T. A. & Drapier, J. C. (2003) Biochemistry42, 7648–7654. [[PubMed]
49. Gunther, M. R., Hsi, L. C., Curtis, J. F., Gierse, J. K., Marnett, L. J., Eling, T. E. & Mason, R. P. (1997) J. Biol. Chem.272, 17086–17090. [[PubMed]
50. Esteves, P. M., De M Carneiro, J. W., Cardoso, S. P., Barbosa, A. G., Laali, K. K., Rasul, G., Prakash, G. K. & Olah, G. A. (2003) J. Am. Chem. Soc.125, 4836–4849. [[PubMed]
51. Quijano, C., Hernandez-Saavedra, D., Castro, L., McCord, J. M., Freeman, B. A. & Radi, R. (2001) J. Biol. Chem.276, 11631–11638. [[PubMed]
52. Whiteman, M., Siau, J. L. & Halliwell, B. (2003) J. Biol. Chem.278, 8380–8384. [[PubMed]
53. Aslan, M., Ryan, T. M., Townes, T. M., Coward, L., Kirk, M. C., Barnes, S., Alexander, C. B., Rosenfeld, S. S. & Freeman, B. A. (2003) J. Biol. Chem.278, 4194–4204. [[PubMed]
54. Grune, T., Blasig, I. E., Sitte, N., Roloff, B., Haseloff, R. & Davies, K. J. (1998) J. Biol. Chem.273, 10857–10862. [[PubMed]
55. Irie, Y., Saeki, M., Kamisaki, Y., Martin, E. & Murad, F. (2003) Proc. Natl. Acad. Sci. USA100, 5634–5639.
56. Rubbo, H., Radi, R., Trujillo, M., Telleri, R., Kalyanaraman, B., Barnes, S., Kirk, M. & Freeman, B. A. (1994) J. Biol. Chem.269, 26066–26075. [[PubMed]
57. Miles, A. M., Bohle, D. S., Glassbrenner, P. A., Hansert, B., Wink, D. A. & Grisham, M. B. (1996) J. Biol. Chem.271, 40–47. [[PubMed]
58. Goldstein, S., Czapski, G., Lind, J. & Merenyi, G. (2000) J. Biol. Chem.275, 3031–3036. [[PubMed]
59. Estevez, A. G., Crow, J. P., Sampson, J. B., Reiter, C., Zhuang, Y., Richardson, G. J., Tarpey, M. M., Barbeito, L. & Beckman, J. S. (1999) Science286, 2498–2500. [[PubMed]
60. Fries, D. M., Paxinou, E., Themistocleous, M., Swanberg, E., Griendling, K. K., Salvemini, D., Slot, J. W., Heijnen, H. F., Hazen, S. L. & Ischiropoulos, H. (2003) J. Biol. Chem.278, 22901–22907. [[PubMed]
61. Jiang, Q. & Hurst, J. K. (1997) J. Biol. Chem.272, 32767–32772. [[PubMed]
62. Rosen, H., Crowley, J. R. & Heinecke, J. W. (2002) J. Biol. Chem.277, 30463–30468. [[PubMed]
63. Baldus, S., Eiserich, J. P., Brennan, M. L., Jackson, R. M., Alexander, C. B. & Freeman, B. A. (2002) Free Radical Biol. Med.33, 1010. [[PubMed]
64. Hazen, S. L., Zhang, R., Shen, Z., Wu, W., Podrez, E. A., MacPherson, J. C., Schmitt, D., Mitra, S. N., Mukhopadhyay, C., Chen, Y., et al. (1999) Circ. Res.85, 950–958. [[PubMed]
65. Souza, J. M., Daikhin, E., Yudkoff, M., Raman, C. S. & Ischiropoulos, H. (1999) Arch. Biochem. Biophys.371, 169–178. [[PubMed]
66. Cassina, A., Hodara, R., Souza, J., Thomson, L., Castro, L., Ischiropoulos, H., Freeman, B. A. & Radi, R. (2000) J. Biol. Chem.275, 21409–21415. [[PubMed]
67. Vadseth, C., Souza, J. M., Thomson, L., Seagraves, A., Nagaswami, C., Scheiner, T., Torbet, J., Vilaire, G., Bennett, J. S., Murciano, J. C., et al. (2003) J. Biol. Chem., jbc/M306101200v1. [[PubMed]
68. Balafanova, Z., Bolli, R., Zhang, J., Zheng, Y., Pass, J. M., Bhatnagar, A., Tang, X.-L., Wang, O., Cardwell, E. & Ping, P. (2002) J. Biol. Chem.277, 15021–15027. [[PubMed]
69. MacMillan-Crow, L. A., Crow, J. P., Kerby, J. D., Beckman, J. S. & Thompson, J. A. (1996) Proc. Natl. Acad. Sci. USA93, 11853–11858.
70. Castro, L., Eiserich, J. P., Sweeney, S., Radi, R. & Freeman, B. A. (2004) Arch. Biochem. Biophys.421, 99–107. [[PubMed]
71. Guo, W., Adachi, T., Matsui, R., Xu, S., Jiang, B., Zou, M. H., Kirber, M., Lieberthal, W. & Cohen, R. A. (2003) Am. J. Physiol.285, H1396–H1403. [[PubMed]
72. Schmidt, P., Youhnovski, N., Daiber, A., Balan, A., Arsic, M., Bachschmid, M., Przybylski, M. & Ullrich, V. (2003) J. Biol. Chem.278, 12813–12819. [[PubMed]
73. Bachschmid, M., Thurau, S., Zou, M. H. & Ullrich, V. (2003) FASEB J.17, 914–916. [[PubMed]
74. Stuehr, D., Pou, S. & Rosen, G. M. (2001) J. Biol. Chem.276, 14533–14536. [[PubMed]
75. Shishehbor, M. H., Aviles, R. J., Brennan, M. L., Fu, X., Goormastic, M., Pearce, G. L., Gokce, N., Keaney, J. F., Jr., Penn, M. S., Sprecher, D. L., et al. (2003) J. Am. Med. Assoc.289, 1675–1680. [[PubMed]
76. Brennan, M. L., Penn, M. S., Van Lente, F., Nambi, V., Shishehbor, M. H., Aviles, R. J., Goormastic, M., Pepoy, M. L., McErlean, E. S., Topol, E. J., et al. (2003) N. Engl. J. Med.349, 1595–1604. [[PubMed]
77. Radi, R., Cassina, A., Hodara, R., Quijano, C. & Castro, L. (2002) Free Radical Biol. Med.33, 1451–1464. [[PubMed]
78. Turko, I. V., Li, L., Aulak, K. S., Stuehr, D. J., Chang, J. Y. & Murad, F. (2003) J. Biol. Chem.278, 33972–33977. [[PubMed]
79. Pearce, L. L., Pitt, B. R. & Peterson, J. (1999) J. Biol. Chem.274, 35763–35767. [[PubMed]
80. Pietraforte, D., Salzano, A. M., Scorza, G., Marino, G. & Minetti, M. (2001) Biochemistry40, 15300–15309. [[PubMed]
81. Herold, S., Shivashankar, K. & Mehl, M. (2002) Biochemistry41, 13460–13472. [[PubMed]
82. Romero, N., Radi, R., Linares, E., Augusto, O., Detweiler, C. D., Mason, R. P. & Denicola, A. (2003) J. Biol. Chem.278, 44049–44057. [[PubMed]
83. Romero, N., Denicola, A., Souza, J. M. & Radi, R. (1999) Arch. Biochem. Biophys.368, 23–30. [[PubMed]
84. Cosby, K., Partovi, K. S., Crawford, J. H., Patel, R. P., Reiter, C. D., Martyr, S., Yang, B. K., Waclawiw, M. A., Zalos, G., Xu, X., et al. (2003) Nat. Med.9, 1498–1505. [[PubMed]
85. Estevez, A. G., Sampson, J. B., Zhuang, Y., Spear, N., Richardson, G. J., Crow, J. P., Tarpey, M. M., Barbeito, L. & Beckman, J. S. (2000) Free Radical Biol. Med.28, 437–446. [[PubMed]
86. Bartesaghi, S., Trujillo, M., Denicola, A., Folkes, L., Wardman, P. & Radi, R. (2004) Free Radical Biol. Med.36, 471–483. [[PubMed]
87. Scott, G. S. & Hooper, D. C. (2001) Med. Hypotheses56, 95–100. [[PubMed]
88. Trujillo, M., Alvarez, M. N., Peluffo, G., Freeman, B. A. & Radi, R. (1998) J. Biol. Chem.273, 7828–7834. [[PubMed]
89. Ferrer-Sueta, G., Vitturi, D., Batinic-Haberle, I., Fridovich, I., Goldstein, S., Czapski, G. & Radi, R. (2003) J. Biol. Chem.278, 27432–27438. [[PubMed]
90. Muscoli, C., Cuzzocrea, S., Riley, D. P., Zweier, J. L., Thiemermann, C., Wang, Z. Q. & Salvemini, D. (2003) Br. J. Pharmacol.140, 445–460.
91. Pacher, P., Liaudet, L., Bai, P., Mabley, J. G., Kaminski, P. M., Virag, L., Deb, A., Szabo, E., Ungvari, Z., Wolin, M. S., Groves, J. T. & Szabo, C. (2003) Circulation107, 896–904. [[PubMed]
92. Keller, J. N., Kindy, M. S., Holtsberg, F. W., St Clair, D. K., Yen, H. C., Germeyer, A., Steiner, S. M., Bruce-Keller, A. J., Hutchins, J. B. & Mattson, M. P. (1998) J. Neurosci.18, 687–697.
93. Wang, H. D., Johns, D. G., Xu, S. & Cohen, R. A. (2002) Am. J. Physiol.282, H1697–H1702. [[PubMed]
94. Kim, G. W. & Chan, P. H. J (2002) Cereb. Blood Flow Metab.22, 798–809. [[PubMed]
95. Gaut, J. P., Byun, J., Tran, H. D., Lauber, W. M., Carroll, J. A., Hotchkiss, R. S., Belaaouaj, A. & Heinecke, J. W. (2002) J. Clin. Invest.109, 1311–1319.
96. Brennan, M. L. Anderson, M. M., Shih, D. M., Qu, X. D., Wang, X., Mehta, A. C., Lim, L. L., Shi, W., Hazen, S. L., Jacob, J. S., et al. (2001) J. Clin. Invest.107, 419–430.
97. LaVaute, T., Smith, S., Cooperman, S., Iwai, K., Land, W., Meyron-Holtz, E., Drake, SK., Miller, G., Abu-Asab, M., Tsokos, M., et al. (2001) Nat. Genet.27, 209–214. [[PubMed][Google Scholar]