Measurement of Nitric Oxide Levels in the Red Cell

Wang, Xunde; Bryan, Nathan S.; MacArthur, Peter H.; Rodríguez, Juan; Gladwin, Mark T.; Feelisch, Martin

doi:10.1074/jbc.m603953200

Cited by 78 publications

(57 citation statements)

References 42 publications

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“…As an initial validation, we synthesized stock solutions of SNO oxyHb (50-100 M SNO/100 M Hb as assayed by photolysis; Ϸ15% Met) and verified under these conditions that amounts of SNO when measured by photolysis, triiodide, and Saville assays (39) were in good agreement (values by triiodide and photolysis were within 5%, P was not significant). As previously described, stock solutions were relatively resistant to SAA in the triiodide assay (46). However, upon dilution of the stock directly into Hb (100 M) (1:10 to 1:1,000) the SNO became progressively sensitive to SAA; at a ratio of 1:1,000 (100 nM SNO-Hb/100 M Hb), simulating RBCs (see Fig.…”

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 94%

“…SNO-Protein Stability. It has been asserted that SNOs and FeNOs in general and SNO-Hbs in particular are stable in acid (46). However, it is well know that acid can destabilize metal nitrosyls (17,58) and SNOs (Eqs.…”

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

“…The triiodide assay has also been used to differentiate SNO-Hb and Hb [FeNO] in vivo on the basis of reactivity toward FeCN/KCN (41)(42)(43)(44)(45)(46). We tested the validity of this approach by analysis of another SNO-Hb redox valency hybrid containing Fe(III) (synthesized with NO solution by using method 1).…”

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

“…The reactivity of S-nitrosylated Hb depends on the redox valency state, the ligation state of the hemes, and the number of SNO/Hb (5,12,37,(47)(48)(49). SNO-Hb standards previously used in triiodide assays (40)(41)(42)(43)(44)(45)(46) were synthesized with a large excess of low-molecular-weight SNO, which produces SNO-Hbs with Ϸ2 SNOs per tetramer; heme valency and ligation states of the standards were not characterized, and the samples were not assayed at physiological ratios of SNO/Hb. As an initial validation, we synthesized stock solutions of SNO oxyHb (50-100 M SNO/100 M Hb as assayed by photolysis; Ϸ15% Met) and verified under these conditions that amounts of SNO when measured by photolysis, triiodide, and Saville assays (39) were in good agreement (values by triiodide and photolysis were within 5%, P was not significant).…”

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

“…Moreover, few NO standards have actually been tested, and no basis has been provided for asserting that response of these standards captures the general behavior. Recovery of certain FeNO standards is reported to be as low as zero (40), and the one SNO-Hb standard that has been widely used (an R-structured Hb that contains Ϸ2 NO per tetramer) (40)(41)(42)(43)(44)(45)(46) is neither characteristic of general SNO-Hb reactivity nor of the reactivity of the micropopulation found in RBCs (a valency hybrid, estimated 1 NO per tetramer) (5,47). Furthermore, the claimed effects of added reagents in triiodide assays, including FeCN and NEM (alone and in combination), are not supported, and they, along with the acidic and denaturing conditions of the triiodide assay, can alter the reactivities of SNO and FeNO as well as disrupt the partitioning of NO species within hydrophobic compartments and thus lead to their misidentification.…”

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confidence: 99%

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Assessment of nitric oxide signals by triiodide chemiluminescence

Hausladen

Rafikov

Angelo

et al. 2007

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

101

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Nitric oxide (NO) bioactivity is mainly conveyed through reactions with iron and thiols, furnishing iron nitrosyls and S-nitrosothiols with wide-ranging stabilities and reactivities. Triiodide chemiluminescence methodology has been popularized as uniquely capable of quantifying these species together with NO byproducts, such as nitrite and nitrosamines. Studies with triiodide, however, have challenged basic ideas of NO biochemistry. The assay, which involves addition of multiple reagents whose chemistry is not fully understood, thus requires extensive validation: Few protein standards have in fact been characterized; NO mass balance in biological mixtures has not been verified; and recovery of species that span the range of NO-group reactivities has not been assessed. Here we report on the performance of the triiodide assay vs. photolysis chemiluminescence in side-by-side assays of multiple nitrosylated standards of varied reactivities and in assays of endogenous Fe-and S-nitrosylated hemoglobin. Although the photolysis method consistently gives quantitative recoveries, the yields by triiodide are variable and generally low (approaching zero with some standards and endogenous samples). Moreover, in triiodide, added chemical reagents, changes in sample pH, and altered ionic composition result in decreased recoveries and misidentification of NO species. We further show that triiodide, rather than directly and exclusively producing NO, also produces the highly potent nitrosating agent, nitrosyliodide. Overall, we find that the triiodide assay is strongly influenced by sample composition and reactivity and does not reliably identify, quantify, or differentiate NO species in complex biological mixtures. red blood cell vasodilation ͉ S-nitrosohemoglobin ͉ S-nitrosylationT he biological effects of nitric oxide (NO) are mediated in large part through binding to transition metals and cysteine thiols at active or allosteric sites within regulatory proteins (1), which elicits changes in protein activity, protein-protein interactions, and protein location (1). Within tissues, many dozens of S-nitrosylated proteins have been identified, and signatures of NO bound to nonheme and heme iron have been detected (1, 2). Additionally, NO can be transported in endocrine or paracrine fashion by reacting with heme iron and cysteine thiols in proteins [hemoglobin (Hb) and albumin] and peptides (glutathione and cysteinlyglycine) to form NO adducts with longer biological lifetimes (3-5); release of NO bioactivity from stable adducts is effected by allosteric and redox-based mechanisms that alter FeNO or S-nitrosothiol (SNO) reactivity (5, 6). An updated discussion of the factors influencing reactivity of S-nitrosohemoglobin, S-nitrosoalbumin, and low-molecular-weight SNO in the context of vasoregulation (5-15) can be found in supporting information (SI) Text.The dynamic distribution of protein and low-molecular-weight NO compounds that subserve NO transport and signaling instantiate the variation in both FeNO and SNO reactivities (4,6,13...

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Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 94%

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

Section: See Also Triiodide Cautionary Notes In Si Text)mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Assessment of nitric oxide signals by triiodide chemiluminescence

Hausladen

Rafikov

Angelo

et al. 2007

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

101

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Effect of Oral Ranitidine on Urinary Excretion ofN-Nitrosodimethylamine (NDMA)

Florian

Matta

DePalma

et al. 2021

JAMA

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IMPORTANCE In 2019, the US Food and Drug Administration (FDA) received a citizen petition indicating that ranitidine contained the probable human carcinogen N-nitrosodimethylamine (NDMA). In addition, the petitioner proposed that ranitidine could convert to NDMA in humans; however, this was primarily based on a small clinical study that detected an increase in urinary excretion of NDMA after oral ranitidine consumption.OBJECTIVE To evaluate the 24-hour urinary excretion of NDMA after oral administration of ranitidine compared with placebo. DESIGN, SETTING, AND PARTICIPANTS Randomized, double-blind, placebo-controlled, crossover clinical trial at a clinical pharmacology unit (West Bend, Wisconsin) conducted in 18 healthy participants. The study began in June 2020, and the end of participant follow-up was July 1, 2020.INTERVENTIONS Participants were randomized to 1 of 4 treatment sequences and over 4 periods received ranitidine (300 mg) and placebo (randomized order) with a noncured-meats diet and then a cured-meats diet. The cured-meats diet was designed to have higher nitrites, nitrates (nitrate-reducing bacteria can convert nitrates to nitrites), and NDMA. MAIN OUTCOME AND MEASURE Twenty-four-hour urinary excretion of NDMA.RESULTS Among 18 randomized participants (median age, 33.0 [interquartile range {IQR}, 28.3 to 42.8] years; 9 women [50%]; 7 White [39%], 11 African American [61%]; and 3 Hispanic or Latino ethnicity [17%]), 17 (94%) completed the trial. The median 24-hour NDMA urinary excretion values for ranitidine and placebo were 0.6 ng (IQR, 0 to 29.7) and 10.5 ng (IQR, 0 to 17.8), respectively, with a noncured-meats diet and 11.9 ng (IQR, 5.6 to 48.6) and 23.4 ng (IQR, 8.6 to 36.7), respectively, with a cured-meats diet. There was no statistically significant difference between ranitidine and placebo in 24-hour urinary excretion of NDMA with a noncured-meats diet (median of the paired differences, 0 [IQR, −6.9 to 0] ng; P = .54) or a cured-meats diet (median of the paired differences, −1.1 [IQR, −9.1 to 11.5] ng; P = .71). No drug-related serious adverse events were reported. CONCLUSIONS AND RELEVANCEIn this trial that included 18 healthy participants, oral ranitidine (300 mg), compared with placebo, did not significantly increase 24-hour urinary excretion of NDMA when participants consumed noncured-meats or cured-meats diets. The findings do not support that ranitidine is converted to NDMA in a general, healthy population.

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Nitric oxide‐releasing polyurethane–PEG copolymer containing the YIGSR peptide promotes endothelialization with decreased platelet adhesion

et al. 2007

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Thrombosis and intimal hyperplasia are the principal causes of small-diameter vascular graft failure. To improve the long-term patency of polyurethane vascular grafts, we have incorporated both poly(ethylene glycol) and a diazeniumdiolate nitric oxide (NO) donor into the backbone of polyurethane to improve thromboresistance. Additionally, we have incorporated the laminin-derived cell adhesive peptide sequence YIGSR to encourage endothelial cell adhesion and migration, while NO release encourages endothelial cell proliferation. NO production by polyurethane films under physiological conditions demonstrated biphasic release, in which an initial burst of 70% of the incorporated NO was released within 2 days, followed by sustained release over 2 months. Endothelial cell proliferation in the presence of the NO-releasing material was increased as compared to control polyurethane, and platelet adhesion to polyethylene glycol-containing polyurethane was decreased significantly with the addition of the NO donor.

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Measurement of Nitric Oxide Levels in the Red Cell

Cited by 78 publications

References 42 publications

Assessment of nitric oxide signals by triiodide chemiluminescence

Assessment of nitric oxide signals by triiodide chemiluminescence

Effect of Oral Ranitidine on Urinary Excretion ofN-Nitrosodimethylamine (NDMA)

Nitric oxide‐releasing polyurethane–PEG copolymer containing the YIGSR peptide promotes endothelialization with decreased platelet adhesion

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