UPLC–MS/MS measurement of S-nitrosoglutathione (GSNO) in human plasma solves the S-nitrosothiol concentration enigma

Tsikas, Dimitrios; Schmidt, Mario; Böhmer, Anke; Zoerner, Alexander A.; Gutzki, Frank‐Mathias; Jordan, Jens

doi:10.1016/j.jchromb.2013.01.023

Cited by 41 publications

(24 citation statements)

References 43 publications

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“…However, there are still many challenges with purification and functional characterization of nitrated proteins. Although we and other laboratories reported many nitrated proteins [35, 47, 179, 189–200], and Table 1 it is conceivable that the actual nitrated proteins should be far greater than those we identified in APAP-exposed WT mice. Some nitrated proteins are transiently expressed while the amounts of other nitrated proteins could be decreased possibly through proteolytic degradation upon nitration, as previously mentioned.…”

Section: Detection Methods Of Nitrated Proteins and Challengescontrasting

confidence: 49%

“…Despite the fact that all techniques have merits, there are many short outcomes that can be improved especially when it comes to the number of identified proteins and the ability to capture proteins that is not highly expressed or the capturing of nonspecific proteins. Many excellent reviews have been already reported with more details about nitrated protein isolation and characterization protocols and the advantages and disadvantages of each method ([51, 199, 200] and references within).…”

Section: Detection Methods Of Nitrated Proteins and Challengesmentioning

confidence: 99%

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Functional Roles of Protein Nitration in Acute and Chronic Liver Diseases

Abdelmegeed

Song

2014

Oxidative Medicine and Cellular Longevity

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Nitric oxide, when combined with superoxide, produces peroxynitrite, which is known to be an important mediator for a number of diseases including various liver diseases. Peroxynitrite can modify tyrosine residue(s) of many proteins resulting in protein nitration, which may alter structure and function of each target protein. Various proteomics and immunological methods including mass spectrometry combined with both high pressure liquid chromatography and 2D PAGE have been employed to identify and characterize nitrated proteins from pathological tissue samples to determine their roles. However, these methods contain a few technical problems such as low efficiencies with the detection of a limited number of nitrated proteins and labor intensiveness. Therefore, a systematic approach to efficiently identify nitrated proteins and characterize their functional roles is likely to shed new insights into understanding of the mechanisms of hepatic disease pathophysiology and subsequent development of new therapeutics. The aims of this review are to briefly describe the mechanisms of hepatic diseases. In addition, we specifically describe a systematic approach to efficiently identify nitrated proteins to study their causal roles or functional consequences in promoting acute and chronic liver diseases including alcoholic and nonalcoholic fatty liver diseases. We finally discuss translational research applications by analyzing nitrated proteins in evaluating the efficacies of potentially beneficial agents to prevent or treat various diseases in the liver and other tissues.

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Section: Detection Methods Of Nitrated Proteins and Challengescontrasting

confidence: 49%

Section: Detection Methods Of Nitrated Proteins and Challengesmentioning

confidence: 99%

Functional Roles of Protein Nitration in Acute and Chronic Liver Diseases

Abdelmegeed

Song

2014

Oxidative Medicine and Cellular Longevity

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“…Given that circulating concentrations of SNOs are typically measured to be < 50 nmol/L[42, 43], this result suggests that the sensitivity of the hind limb vessels to L-cysNO is low, and that it is not a physiological regulator of femoral vascular tone. However, systemic MAP was decreased by ~40% with GSNO infusions of 0.4 ml/min, corresponding to estimated plasma SNO concentration of <300 nmol/L (50% of the 600 nmol/L measured at the 0.8 ml/min infusion rate).…”

Section: Discussionmentioning

confidence: 99%

Local and systemic vasodilatory effects of low molecular weight S-nitrosothiols

Liu

Schroeder

Wilson

et al. 2016

Free Radical Biology and Medicine

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S-nitrosothiols (SNOs) such as S-nitroso-L-cysteine (L-cysNO) are endogenous compounds with potent vasodilatory activity. During circulation in the blood, the NO moiety can be exchanged among various thiol-containing compounds by S-transnitrosylation, resulting in SNOs with differing capacities to enter the cell (membrane permeability). To determine whether the vasodilating potency of SNOs is dependent upon membrane permeability, membrane-permeable L-cysNO and impermeable S-nitroso-D-cysteine (D-cysNO) and S-nitroso-glutathione (GSNO) were infused into one femoral artery of anesthetized adult sheep while measuring bilateral femoral and systemic vascular conductances. L-cysNO induced vasodilation in the infused hind limb, whereas D-cysNO and GSNO did not. L-cysNO also increased intracellular NO in isolated arterial smooth muscle cells, whereas GSNO did not. The infused SNOs remained predominantly in a low molecular weight form during first-passage through the hind limb vasculature, but were converted into high molecular weight SNOs upon systemic recirculation. At systemic concentrations of ~0.6 μmol/L, all three SNOs reduced mean arterial blood pressure by ~50%, with pronounced vasodilation in the mesenteric bed. Pharmacokinetics of L-cysNO and GSNO were measured in vitro and in vivo and correlated with their hemodynamic effects, membrane permeability, and S-transnitrosylation. These results suggest local vasodilation by SNOs in the hind limb requires membrane permeation, whereas systemic vasodilation does not. The systemic hemodynamic effects of SNOs occur after equilibration of the NO moiety amongst the plasma thiols via S-transnitrosylation.

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“…In fact, use of NEM in the analysis of these molecules both prevents S -transnitrosation reactions with physiological thiols and avoids artifactual increase of RSNO concentration during the acidification step, when nitrite is converted into nitrous acid, a strong nitrosating agent [29]. However, we did not include the measurement of these molecules in our protocol because of their extremely low concentration, that makes their analysis very time consuming and dependent on expensive instrumentation [29,30]. …”

Section: Discussionmentioning

confidence: 99%

Immediate stabilization of human blood for delayed quantification of endogenous thiols and disulfides

Giustarini

Galvagni

Orlandini

et al. 2016

Journal of Chromatography B

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Endogenous thiols undergo rapid and reversible oxidation to disulfides when exposed to oxidants and are, therefore, suitable biomarkers of oxidative stress. However, accurate analysis of thiols in blood is frequently compromised by their artifactual oxidation during sample manipulation, which spuriously elevates the disulfide levels. Here, we describe a validated pre-analytical procedure that prevents both artifactual oxidation of thiols during sample manipulation and their oxidative decay for months in biosamples that are stored at −80°C. Addition of N-ethylmaleimide to blood samples from healthy donors was used to stabilize whole blood, red blood cells, platelets and plasma disulfides, whereas addition of citrate buffer followed by dilution of plasma with H2O was used to stabilize plasma thiols. The concentrations of thiols and disulfides were stable in all biosamples for at least 6 months when analyzed by UV/Vis HPLC at regular intervals. Only 3 ml of blood were needed to perform the analyses of thiols and disulfides in the different blood fractions. This pre-analytical procedure is reliable for use in both animal and human prospective studies. Its ease of implementation makes the method suitable for application to multicenter studies where blood samples are collected by different sites and personnel and are shipped to specific specialized laboratories.

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UPLC–MS/MS measurement of S-nitrosoglutathione (GSNO) in human plasma solves the S-nitrosothiol concentration enigma

Cited by 41 publications

References 43 publications

Functional Roles of Protein Nitration in Acute and Chronic Liver Diseases

Functional Roles of Protein Nitration in Acute and Chronic Liver Diseases

Local and systemic vasodilatory effects of low molecular weight S-nitrosothiols

Immediate stabilization of human blood for delayed quantification of endogenous thiols and disulfides

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