[19] Antibodies that recognize nitrotyrosine

Ye, Yao Zu; Strong, Michael J.; Huang, Zhiqiang; Beckman, Joseph S.

doi:10.1016/s0076-6879(96)69022-3

Cited by 184 publications

(113 citation statements)

References 10 publications

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“…Dot blot quantitation of nitrotyrosine in motor neurons was performed by harvesting total protein from 30,000 motor neurons plated in a 35-mm dish for 24 h. Samples containing equal amounts of protein were blotted on a nitrocellulose membrane by gravity flow using a Bio-Blot microfiltration apparatus (Bio-Rad). Membranes were then processed as described for Western blot using rabbit polyclonal primary antibody to nitrotyrosine as described previously (70). Immunoreactivity was visualized with SuperSignal Western blot kit (Pierce) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Prevention of Peroxynitrite-induced Apoptosis of Motor Neurons and PC12 Cells by Tyrosine-containing Peptides

Quijano

Robinson

et al. 2007

Journal of Biological Chemistry

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Although peroxynitrite stimulates apoptosis in many cell types, whether peroxynitrite acts directly as an oxidant or the induction of apoptosis is because of the radicals derived from peroxynitrite decomposition remains unknown. Before undergoing apoptosis because of trophic factor deprivation, primary motor neuron cultures become immunoreactive for nitrotyrosine. We show here using tyrosine-containing peptides that free radical processes mediated by peroxynitrite decomposition products were required for triggering apoptosis in primary motor neurons and in PC12 cells cultures. The same concentrations of tyrosine-containing peptides required to prevent the nitration and apoptosis of motor neurons induced by trophic factor deprivation and of PC12 cells induced by peroxynitrite also prevented peroxynitrite-mediated nitration of motor neurons, brain homogenates, and PC12 cells. The heat shock protein 90 chaperone was nitrated in both trophic factor-deprived motor neurons and PC12 cells incubated with peroxynitrite. Tyrosine-containing peptides did not affect the induction of PC12 cell death by hydrogen peroxide. Tyrosine-containing peptides should protect by scavenging peroxynitrite-derived radicals and not by direct reactions with peroxynitrite as they neither increase the rate of peroxynitrite decomposition nor decrease the bimolecular peroxynitrite-mediated oxidation of thiols. These results reveal an important role for free radicalmediated nitration of tyrosine residues, in apoptosis induced by endogenously produced and exogenously added peroxynitrite; moreover, tyrosine-containing peptides may offer a novel strategy to neutralize the toxic effects of peroxynitrite.

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Section: Methodsmentioning

confidence: 99%

Prevention of Peroxynitrite-induced Apoptosis of Motor Neurons and PC12 Cells by Tyrosine-containing Peptides

Quijano

Robinson

et al. 2007

Journal of Biological Chemistry

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“…The sensitivity was 25 pg?mL -1 . Secondly, nNOS (NOS1) and iNOS (NOS2) were assessed using standard ELISA sandwich techniques, as described previously [18]. For the detection of nNOS, the monoclonal and polyclonal antibodies were purchased from BD Biosciences (San Diego, CA, USA) and the recombinant enzyme for the standard curve was purchased from Calbiochem (San Diego, CA, USA).…”

Section: Muscle Biopsymentioning

confidence: 99%

Skeletal muscle inflammation and nitric oxide in patients with COPD

Torres²,

Sanctis³

et al. 2005

Eur Respir J

105

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In chronic obstructive pulmonary disease (COPD) the presence of systemic inflammation has been associated with peripheral muscle abnormalities and weight loss.To study whether inflammatory factors are important in these processes, the present study compared the skeletal muscle levels of nitrite, nitrate, nitrotyrosine, neuronal, endothelial and inducible nitric oxide synthases (nNOS, eNOS, and iNOS, respectively), and inflammatory markers (tumour necrosis factor (TNF)-a, CD154 and CD163) in 15 patients (forced expiratory volume in one second 43¡11%) and 14 controls. All these markers were also compared between patients with normal and low body weight.Nitrite ( controls. Similar levels of all these markers were observed in COPD patients with normal and low body weight.In conclusion, these findings suggest the presence of an inflammatory process in the muscle tissue of chronic obstructive pulmonary disease patients, and argue in favour of its participation in the pathogenesis of skeletal muscle abnormalities.

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“…Proteins were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis using 12% gels and transferred to nitrocellulose. The membranes were incubated with monoclonal 1A6 anti-nitrotyrosine [1:500 (Beckman et al,l994b)], washed, probed with biotinylated anti-mouse IgG, and developed using avidin-horseradish peroxidase plus diaminobenzidine (Ye et al, 1996).…”

Section: Nitration In Vitromentioning

confidence: 99%

Superoxide Dismutase Catalyzes Nitration of Tyrosines by Peroxynitrite in the Rod and Head Domains of Neurofilament‐L

Crow

Ye²,

Strong

et al. 1997

Journal of Neurochemistry

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236

123

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Superoxide dismutase (SOD) catalyzes the nitration of specific tyrosine residues in proteins by peroxynitrite (ONOO), which may be the damaging gain-offunction resulting from mutations to SOD associated with familial amyotrophic lateral sclerosis (ALS). We found that disassembled neurofilament-L (light subunit) was more susceptible to tyrosine nitration catalyzed by SOD in vitro. Neurofilament-L was selectively nitrated compared with the majority of other proteins present in brain homogenates. Assembled neurofilament-L was more resistant to nitration, suggesting that the susceptible tyrosine residues were protected by intersubunit contacts in assembled neurofilaments. Electrospray mass spectrometry of trypsin-digested neurofilament-L showed that tyrosine 17 in the head region and tyrosines 138, 177, and 265 in a-helical coil regions of the rod domain of neurofilament-L were particularly susceptible to SOD-catalyzed nitration. Nitrated neurofilament-L inhibited the assembly of unmodified neurofilament subunits, suggesting that the affected tyrosines are located in regions important for intersubunit contacts. Neurofilaments are major structural proteins expressed in motor neurons and known to be important for their survival in vivo. We suggest that SODcatalyzed nitration of neurofilament-L may have a significant role in the pathogenesis of ALS.

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[19] Antibodies that recognize nitrotyrosine

Cited by 184 publications

References 10 publications

Prevention of Peroxynitrite-induced Apoptosis of Motor Neurons and PC12 Cells by Tyrosine-containing Peptides

Prevention of Peroxynitrite-induced Apoptosis of Motor Neurons and PC12 Cells by Tyrosine-containing Peptides

Skeletal muscle inflammation and nitric oxide in patients with COPD

Superoxide Dismutase Catalyzes Nitration of Tyrosines by Peroxynitrite in the Rod and Head Domains of Neurofilament‐L

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