[21] Detection of S-nitrosothiols by fluorometric and colorimetric methods

Wink, David A.; Kim, Sungmee; Coffin, Deborah; Cook, John C.; Vodovotz, Yoram; Chistodoulou, Danae; Jourd’heuil, David; Grisham, Matthew B.

doi:10.1016/s0076-6879(99)01083-6

Cited by 64 publications

(55 citation statements)

References 26 publications

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“…First, in a specific fluorescence assay for S-nitrosylated proteins, we demonstrated the reaction of recombinant parkin with the physiological NO donor, SNOC. This assay detects the formation of S-nitrosothiol by measurement of the fluorescent compound NAT (14,20). NAT is stoichiometrically converted from 2,3-diaminonaphthalene by NO released from Snitrosylated proteins and thus provided a quantitative measure of S-nitrosothiol formation on parkin.…”

Section: S-nitrosylation Of Parkin In Vitromentioning

confidence: 99%

Nitrosative stress linked to sporadic Parkinson's disease: S-nitrosylation of parkin regulates its E3 ubiquitin ligase activity

Yao

Nakamura

et al. 2004

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

488

464

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Many hereditary and sporadic neurodegenerative disorders are characterized by the accumulation of aberrant proteins. In sporadic Parkinson's disease, representing the most prevalent movement disorder, oxidative and nitrosative stress are believed to contribute to disease pathogenesis, but the exact molecular basis for protein aggregation remains unclear. In the case of autosomal recessive-juvenile Parkinsonism, mutation in the E3 ubiquitin ligase protein parkin is linked to death of dopaminergic neurons. Here we show both in vitro and in vivo that nitrosative stress leads to S-nitrosylation of wild-type parkin and, initially, to a dramatic increase followed by a decrease in the E3 ligase-ubiquitin-proteasome degradative pathway. The initial increase in parkin's E3 ubiquitin ligase activity leads to autoubiquitination of parkin and subsequent inhibition of its activity, which would impair ubiquitination and clearance of parkin substrates. These findings may thus provide a molecular link between free radical toxicity and protein accumulation in sporadic Parkinson's disease

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Section: S-nitrosylation Of Parkin In Vitromentioning

confidence: 99%

Nitrosative stress linked to sporadic Parkinson's disease: S-nitrosylation of parkin regulates its E3 ubiquitin ligase activity

Yao

Nakamura

et al. 2004

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

488

464

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“…in pH 7.4 PB followed by treatment with acidified-cysteine formed a red solution with a characteristic spectral pattern for S-nitrosocysteine (λ max = 542 nm). 37 Chemiluminescence assay also independently confirmed the formation of S-nitrosothiol.…”

Section: Resultsmentioning

confidence: 70%

Allylic Nitro Compounds as Nitrite Donors

2006

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Allylic nitro compounds were synthesized and evaluated as organic sources of nitrite and nitric oxide. Unactivated allylic nitro compounds do not spontaneously release nitrite and nucleophile promoted nitrite release is slow. However, 2-(nitromethyl)-cyclohex-1-ene-3-one spontaneously dissociates in buffer (pH = 7.4) to release nitrite with a k obs = 1.6 × 10 −5 s −1 . In the presence of Lcysteine, this compound rapidly yields nitrite and reacts with hemoglobin similarly to sodium nitrite. Structural modifications and the nature and amount of nucleophile modulate the rate of nitrite release. In the presence of L-cysteine and ascorbic acid, this compound forms nitric oxide. Together, these results reveal a new structural architecture for the tunable liberation of nitrite and nitric oxide from organic compounds.

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“…Generation of SNO-SHP-2 was followed chemically with a modified DAN assay, as we and others have described previously (10,11,37,45). Briefly, recombinant SHP-2 proteins were expressed and purified from Escherichia coli BL21(DE3).…”

Section: Methodsmentioning

confidence: 99%

S-nitrosylated SHP-2 contributes to NMDA receptor-mediated excitotoxicity in acute ischemic stroke

Shi

Sunico

McKercher

et al. 2013

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

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Overproduction of nitric oxide (NO) can cause neuronal damage, contributing to the pathogenesis of several neurodegenerative diseases and stroke (i.e., focal cerebral ischemia). NO can mediate neurotoxic effects at least in part via protein S-nitrosylation, a reaction that covalently attaches NO to a cysteine thiol (or thiolate anion) to form an S-nitrosothiol. Recently, the tyrosine phosphatase Src homology region 2-containing protein tyrosine phosphatase-2 (SHP-2) and its downstream pathways have emerged as important mediators of cell survival. Here we report that in neurons and brain tissue NO can S-nitrosylate SHP-2 at its active site cysteine, forming S-nitrosylated SHP-2 (SNO-SHP-2). We found that NMDA exposure in vitro and transient focal cerebral ischemia in vivo resulted in increased levels of SNO-SHP-2. S-Nitrosylation of SHP-2 inhibited its phosphatase activity, blocking downstream activation of the neuroprotective physiological ERK1/2 pathway, thus increasing susceptibility to NMDA receptor-mediated excitotoxicity. These findings suggest that formation of SNO-SHP-2 represents a key chemical reaction contributing to excitotoxic damage in stroke and potentially other neurological disorders.nitrosative stress | reactive oxygen species | reactive nitrogen species E xcessive activation of NMDA receptors (NMDARs) activates neuronal nitric oxide (NO) synthase (NOS; nNOS) (1) and results in overproduction of NO in the brain, contributing to neuronal damage in a number of neurodegenerative diseases and cerebral ischemia (2, 3). The NO group can react with specific cysteine thiols to regulate protein activity in a process called Snitrosylation (4, 5). As a ubiquitous posttranslational modification, S-nitrosylation has been found to modulate a broad spectrum of proteins affecting neuronal development, synaptic plasticity, protein folding, and cell death (3). Recent discoveries have demonstrated that excessive NO has a negative effect on neuronal survival via S-nitrosylation of a number of neurodegenerative disease-associated proteins, including parkin, protein disulfide isomerase, GAPDH, Cdk5, and dynamin-related protein 1 (6-11).The Src homology-2 domain-containing phosphatase (SHP-2), a member of the ubiquitously expressed protein-tyrosine phosphatase (PTP) family, contains a cysteine residue at its active site (12). SHP-2 is known to localize in the cytosol and nucleus, and plays important biological functions in response to various growth factors, hormones, and cytokines (13). Recent studies have shown that activation of SHP-2 increases survival of various cell types, including neural progenitor cells and neurons, through activation of ERK1/2 (14, 15). SHP-2 is thought to promote ERK signaling by dephosphorylating negative regulators of the Ras-ERK pathway, such as PAG/Cbp, Ras-GAP, or GAP-binding sites on receptor tyrosine kinases or Sprouty proteins (16)(17)(18)(19)(20). Additionally, transient activation of the ERK1/2 signaling cascade has been implicated in regulating neuronal survival after stroke (21-...

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[21] Detection of S-nitrosothiols by fluorometric and colorimetric methods

Cited by 64 publications

References 26 publications

Nitrosative stress linked to sporadic Parkinson's disease: S-nitrosylation of parkin regulates its E3 ubiquitin ligase activity

Nitrosative stress linked to sporadic Parkinson's disease: S-nitrosylation of parkin regulates its E3 ubiquitin ligase activity

Allylic Nitro Compounds as Nitrite Donors

S-nitrosylated SHP-2 contributes to NMDA receptor-mediated excitotoxicity in acute ischemic stroke

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