Characterization of basal nitric oxide production in living cells

López-Figueroa, Manuel O.; Caamaño, Claudio A.; Marín, Raquel; Guerra, Borja; Alonso, Rafael; Morano, M. Inés; Akil, Huda; Watson, Stanley J.

doi:10.1016/s0167-4889(01)00138-0

Cited by 25 publications

(21 citation statements)

References 30 publications

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“…5A). At 1 mM carboxy-PTIO (NO-specific scavenger), NO production was significantly abolished in COS-1 cells (López-Figueroa et al 2001). As shown in Fig.…”

Section: No Involvement In Hwc-induced Nr Transcript Accumulationmentioning

confidence: 62%

Nitrate reductase, a nitric oxide-producing enzyme: induction by pathogen signals

Yamamoto

Katou

Yoshioka

et al. 2003

Journal of General Plant Pathology

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Nitric oxide (NO) is believed to act as an effector for defense signaling in plant cells. Nitrate reductase (NR) is one of the NO-producing enzymes in plants. Here, we report that infection of Phytophthora infestans, the fungal pathogen of potato late blight, into potato tubers caused a transient increase in the NR transcript in an incompatible, but not a compatible, interaction. Treatment of potato tubers with the fungal elicitor hyphal wall components (HWC) from the fungus induced NR gene at the transcriptional and protein level. Soluble proteins from HWCtreated tubers exhibited enhanced nitrite-dependent NO production. The inhibitor experiments for protein kinase and extracellular Ca 2ϩ on the HWC-induced accumulation of the NR transcript suggested involvement of a calciumindependent protein kinase in regulation of the NR gene. Additionally, we isolated cDNA clones encoding NR from a potato tuber cDNA library; two isogenes were designated StNR5 and StNR6, respectively. Reverse transcription-polymerase chain reaction analyses using gene-specific primers showed that transcripts for both StNR5 and StNR6 were induced by HWC in potato tubers. This is the first report that pathogen signals induce expression of the NR gene.

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“…5A). At 1 mM carboxy-PTIO (NO-specific scavenger), NO production was significantly abolished in COS-1 cells (López-Figueroa et al 2001). As shown in Fig.…”

Section: No Involvement In Hwc-induced Nr Transcript Accumulationmentioning

confidence: 62%

Nitrate reductase, a nitric oxide-producing enzyme: induction by pathogen signals

Yamamoto

Katou

Yoshioka

et al. 2003

Journal of General Plant Pathology

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“…1B). To confirm that S-nitrosylation not only occurred through the treatment with an exogenous NO donor, we performed a similar experiment with the use of N2A cells, which have been shown to possess endogenous neuronal NO synthase (nNOS) activity (8). After the biotin switch assay on N2A cells transfected with XIAP, we found that XIAP was S-nitrosylated under basal conditions (Fig.…”

Section: Bir Domains Of Xiap Are S-nitrosylated By Nomentioning

confidence: 87%

S-nitrosylation of XIAP compromises neuronal survival in Parkinson's disease

Tsang

Lee

et al. 2009

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

147

112

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Inhibitors of apoptosis (IAPs) are a family of highly-conserved proteins that regulate cell survival through binding to caspases, the final executioners of apoptosis. X-linked IAP (XIAP) is the most widely expressed IAP and plays an important function in regulating cell survival. XIAP contains 3 baculoviral IAP repeats (BIRs) followed by a RING finger domain at the C terminal. The BIR domains of XIAP possess anticaspase activities, whereas the RING finger domain enables XIAP to function as an E3 ubiquitin ligase in the ubiquitin and proteasomal system. Our previous study showed that parkin, a protein that is important for the survival of dopaminergic neurons in Parkinson's disease (PD), is S-nitrosylated both in vitro and in vivo in PD patients. S-nitrosylation of parkin compromises its ubiquitin E3 ligase activity and its protective function, which suggests that nitrosative stress is an important factor in regulating neuronal survival during the pathogenesis of PD. In this study we show that XIAP is S-nitrosylated in vitro and in vivo in an animal model of PD and in PD patients. Nitric oxide modifies mainly cysteine residues within the BIR domains. In contrast to parkin, S-nitrosylation of XIAP does not affect its E3 ligase activity, but instead directly compromises its anticaspase-3 and antiapoptotic function. Our results confirm that nitrosative stress contributes to PD pathogenesis through the impairment of prosurvival proteins such as parkin and XIAP through different mechanisms, indicating that abnormal S-nitrosylation plays an important role in the process of neurodegeneration.

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“…It has been suggested that the major advantage of DAF-2 fluorescence spectroscopy over other NO sensitive imaging/ detection techniques (such as electron paramagnetic resonance or EPR with NO spin trapping) is the ability to detect the basal NO production from endothelial and neuronal cells, with submicromolar NO concentrations (41,49,50). The intracellular level of ascorbate is as high as 9 mM in human endothelial cells (24) and is in the range of 1-10 mM in neurons (25).…”

Section: Discussionmentioning

confidence: 99%

Interfering with Nitric Oxide Measurements

Zhang¹,

Kim²,

Hatcher³

et al. 2002

Journal of Biological Chemistry

182

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4,5-Diaminofluorescein (DAF-2) is widely used for detection and imaging of NO based on its sensitivity, noncytotoxicity, and specificity. In the presence of oxygen, NO and NO-related reactive nitrogen species nitrosate 4,5-diaminofluorescein to yield the highly fluorescent DAF-2 triazole (DAF-2T). However, as reported here, the DAF-2 reaction to form a fluorescent product is not specific to NO because it reacts with dehydroascorbic acid (DHA) and ascorbic acid (AA) to generate new compounds that have fluorescence emission profiles similar to that of DAF-2T. When DHA is present, the formation of DAF-2T is attenuated because the DHA competes for DAF-2, whereas AA decreases the nitrosation of DAF-2 to a larger extent, possibly because of additional reducing activity that affects the amount of available N 2 O 3 from the NO. The reaction products of DAF-2 with DHA and AA have been characterized using capillary electrophoresis with laser-induced fluorescence detection and electrospray mass spectrometry. The reactions of DAF-2 with DHA and AA are particularly significant because DHA and AA often colocalize with nitric-oxide synthase in the central nervous, cardiovascular, and immune systems, indicating the importance of understanding this chemistry.NO is involved in a variety of important biological functions in the cardiovascular system, the central and peripheral nervous system, the reproductive system, and the immune system (1-6). NO is normally generated via an enzymatically regulated pathway with the conversion of L-arginine to citrulline by a family of at least three distinct nitric-oxide synthase (NOS) 1 enzymes, the neuronal, inducible, and endothelial NOS forms (or NOS-I, -II, and -III) (7-9). Endothelial and neuronal NOS, also named constitutive NOS, are calcium-dependent. Although cells expressing constitutive NOS generally produce small amounts of NO (submicromolar at the cellular level), inducible NOS, which is synthesized in immune-competent cells when stimulated by cytokines, endotoxins, and other biologically active compounds, is calcium-independent and produces NO at higher levels (1-10 M in microphages) (10). Although in situ and immunohistochemical techniques allow one to determine whether NOS is present in a particular tissue, whether the NOS is actively producing NO under specific conditions and the amount of NO produced are important questions to answer to understand the physiological roles of NOS.Direct detection (or imaging) of NO production in a biological system was unsatisfactory until the development of a series of fluorescent indicators for NO, the diaminofluoresceins (DAFs) in 1998 by the Nagano group (11). In the presence of oxygen, NO nitrosates DAFs to produce the highly fluorescent triazofluoresceins. DAFs provide the advantages of sensitivity (detection limits of 5 nM), simple protocols, and noncytotoxicity, and they are also believed to offer high specificity to NO. Since then, an increasing number of researchers have used them for NO detection and NO imaging (10, 12).The specificity...

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Characterization of basal nitric oxide production in living cells

Cited by 25 publications

References 30 publications

Nitrate reductase, a nitric oxide-producing enzyme: induction by pathogen signals

Nitrate reductase, a nitric oxide-producing enzyme: induction by pathogen signals

S-nitrosylation of XIAP compromises neuronal survival in Parkinson's disease

Interfering with Nitric Oxide Measurements

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