Factors Determining the Selectivity of Protein Tyrosine Nitration

Souza, José Maria de; Daikhin, Evgueni; Yudkoff, Marc; Raman, C.S.; Ischiropoulos, Harry

doi:10.1006/abbi.1999.1480

Cited by 306 publications

(307 citation statements)

References 29 publications

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“…However, nitration often occurs at selective tyrosyl residues possibly biased by proximity to glycyl turns and charged residues (e.g., glutamyl; ref. 34), which are qualities manifest in Tyr-66 of GFP (26). Previous studies have shown that fluorescence of GFP-related peptides can be modified by anions including chloride (35).…”

Section: Discussionmentioning

confidence: 98%

Direct real-time evaluation of nitration with green fluorescent protein in solution and within human cells reveals the impact of nitrogen dioxide vs. peroxynitrite mechanisms

Espey

Xavier

Thomas

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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3-Nitrotyrosyl adducts in proteins have been detected in a wide range of diseases. The mechanisms by which reactive nitrogen oxide species may impede protein function through nitration were examined by using a unique model system, which exploits a critical tyrosyl residue in the fluorophoric pocket of recombinant green fluorescent protein (GFP). Exposure of purified GFP suspended in phosphate buffer to synthetic peroxynitrite in either 0.5 or 5 M steps resulted in progressively increased 3-nitrotyrosyl immunoreactivity concomitant with disappearance of intrinsic fluorescence (IC 50 Ϸ 20 M). Fluorescence from an equivalent amount of GFP expressed within intact MCF-7 tumor cells was largely resistant to this bolus treatment (IC 50 > 250 M). The more physiologically relevant conditions of either peroxynitrite infusion (1 M͞min) or de novo formation by simultaneous, equimolar generation of nitric oxide (NO) and superoxide (e.g., 3-morpholinosydnonimine; NONOates plus xanthine oxidase͞hypo-xanthine, menadione, or mitomycin C) were examined. Despite robust oxidation of dihydrorhodamine under each of these conditions, fluorescence decrease of both purified and intracellular GFP was not evident regardless of carbon dioxide presence, suggesting that oxidation and nitration are not necessarily coupled. Alternatively, both extra-and intracellular GFP fluorescence was exquisitely sensitive to nitration produced by heme-peroxidase͞hydrogen peroxidecatalyzed oxidation of nitrite. Formation of nitrogen dioxide (NO 2) during the reaction between NO and the nitroxide 2-phenyl-4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide indicated that NO 2 can enter cells and alter peptide function through tyrosyl nitration. Taken together, these findings exemplified that heme-peroxidasecatalyzed formation of NO 2 may play a pivotal role in inflammatory and chronic disease settings while calling into question the significance of nitration by peroxynitrite.3-nitrotyrosine ͉ myeloperoxidase ͉ nitric oxide ͉ xanthine oxidase ͉ superoxide T he prevalence of increased 3-nitrotyrosine in both acute inflammatory events and numerous chronic diseases suggests that protein nitration may be an important component in pathophysiologic processes (1-7). However, there is disagreement regarding the predominant mechanism by which tyrosyl nitration occurs under these conditions (1-16). The reaction between superoxide (O 2 Ϫ ) and nitric oxide (NO) results in formation of peroxynitrite (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)17). Numerous studies have concluded that this nitrogen oxide mediates the majority of nitration, on the basis of 3-nitrotyrosine formation after bolus application of synthetically produced peroxynitrite. Nitration by this route is strongly influenced by buffer composition, in particular, the concentration of protons, transition metals, and carbon dioxide (CO 2 ; refs. 1, 6, 20-24). Both neutrophils and macrophages have been shown to elicit nitration reactions through myeloperoxidase (MPO) activity (15-18). Catalysis of nitrite oxidation by heme...

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Section: Discussionmentioning

confidence: 98%

Direct real-time evaluation of nitration with green fluorescent protein in solution and within human cells reveals the impact of nitrogen dioxide vs. peroxynitrite mechanisms

Espey

Xavier

Thomas

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…It is noteworthy to indicate, however, that (i) a relatively limited number of proteins are preferential targets of nitration, and (ii) within these proteins, only one or a few specific tyrosines can be nitrated (65); because the species participating in nitration have short diffusion distances and site-specific nitration occurs at metal-or hemeperoxidase-binding sites, nitration reactions can be concentrated on proteins, cell, or tissue compartments as revealed by immunochemical (with antibodies against protein 3-nitrotyrosine) and proteomic-based methods. Thus, although there is a dilutional effect in quantitating total nitrated protein by analytical techniques in tissues, nitration can be focused on specific tyrosines and potentially result in modification, loss, or gain of function.…”

Section: The Biological Significance Of Protein Tyrosine Nitrationmentioning

confidence: 99%

Nitric oxide, oxidants, and protein tyrosine nitration

Radí

2004

Proc. Natl. Acad. Sci. U.S.A.

1,335

1,066

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“…There appears to be some selectivity in the nitration, since not all proteins in a tissue under analysis are nitrated. Souza et al (1999) carefully analysed nitration of three purified proteins under defined conditions and have determined some of the factors governing selectivity of nitration. Their evidence suggested that exposure of tyrosine at the surface of the protein, its location on a loop structure and an adjacent negative charge were some of the determining factors.…”

Section: Protein Tyrosine-nitrationmentioning

confidence: 99%

Biological chemistry of reactive oxygen and nitrogen and radiation-induced signal transduction mechanisms

2003

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In the past few years, nuclear DNA damage-sensing mechanisms activated by ionizing radiation have been identified, including ATM/ATR and the DNA-dependent protein kinase. Less is known about sensing mechanisms for cytoplasmic ionization events and how these events influence nuclear processes. Several studies have demonstrated the importance of cytoplasmic signaling pathways in cytoprotection and mutagenesis. For cytoplasmic signaling, radiation-stimulated reactive oxygen species (ROS) and reactive nitrogen species (RNS) are essential activators of these pathways. This review summarizes recent studies on the chemistry of radiation-induced ROS/ RNS generation and emphasizes interactions between ROS and RNS and the relative roles of cellular ROS/ RNS generators as amplifiers of the initial ionization events. Cellular mechanisms for regulating ROS/RNS levels are discussed. The mechanisms by which cells sense ROS/RNS are examined in terms of how ROS/RNS modify protein structure and function, for example, interactions with metal-thiol clusters, protein tyrosine nitration, protein cysteine oxidation, S-thiolation and Snitrosylation. We propose that radiation-induced ROS are the initiators and that nitric oxide (NO ) or derivatives are the effectors activating these signal transduction pathways. In responding to cellular ionization events, the cell converts an oxidative signal to a nitrosative one because ROS are too reactive and unspecific in their reactions for regulatory purposes and the cell is equipped to precisely modulate NO levels.

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Factors Determining the Selectivity of Protein Tyrosine Nitration

Cited by 306 publications

References 29 publications

Direct real-time evaluation of nitration with green fluorescent protein in solution and within human cells reveals the impact of nitrogen dioxide vs. peroxynitrite mechanisms

Direct real-time evaluation of nitration with green fluorescent protein in solution and within human cells reveals the impact of nitrogen dioxide vs. peroxynitrite mechanisms

Nitric oxide, oxidants, and protein tyrosine nitration

Biological chemistry of reactive oxygen and nitrogen and radiation-induced signal transduction mechanisms

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