Detection of S-Nitrosoglutathione Reductase Activity in Plants

Kubienová, Lucie; Tichá, Tereza; Luhová, Lenka; Petřivalský, Marek

doi:10.1007/978-1-4939-3600-7_15

Cited by 4 publications

(5 citation statements)

References 16 publications

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“…Detection of GSNOR protein was done by immunoblotting as was described by Kubienová et al (2016). Roots were homogenized in 0.1-M Tris-HCl, pH 7.5 with 1-mM EDTA, 2% (w/v) PVPP, 1-mM DTT, 1% (v/v) protease inhibitor cocktail (Sigma-Aldrich), 0.1% (w/v) Triton X-100, 1-mM MgCl 2 , and 10% (w/v) glycerol in an ice bath.…”

Section: Methodsmentioning

confidence: 99%

Canavanine-Induced Decrease in Nitric Oxide Synthesis Alters Activity of Antioxidant System but Does Not Impact S-Nitrosoglutathione Catabolism in Tomato Roots

et al. 2019

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Canavanine (CAN) is a nonproteinogenic amino acid synthesized in legumes. In mammalians, as arginine analogue, it is an inhibitor of nitric oxide synthase (NOS) activity. The aim of this study was to investigate the impact of CAN-induced nitric oxide level limitation on the antioxidant system and S-nitrosoglutathione (GSNO) metabolism in roots of tomato seedlings. Treatment with CAN (10 or 50 µM) for 24–72 h led to restriction in root growth. Arginine-dependent NOS-like activity was almost completely inhibited, demonstrating direct effect of CAN action. CAN increased total antioxidant capacity and the level of sulphydryl groups. Catalase (CAT) and superoxide dismutase (SOD) activity decreased in CAN exposed roots. CAN supplementation resulted in the decrease of transcript levels of genes coding CAT (with the exception of CAT1). Genes coding SOD (except MnSOD and CuSOD) were upregulated by CAN short treatment; prolonged exposition to 50-µM CAN resulted in downregulation of FeSOD, CuSOD, and SODP-2. Activity of glutathione reductase dropped down after short-term (10-µM CAN) supplementation, while glutathione peroxidase activity was not affected. Transcript levels of glutathione reductase genes declined in response to CAN. Genes coding glutathione peroxidase were upregulated by 50-µM CAN, while 10-µM CAN downregulated GSHPx1. Inhibition of NOS-like activity by CAN resulted in lower GSNO accumulation in root tips. Activity of GSNO reductase was decreased by short-term supplementation with CAN. In contrast, GSNO reductase protein abundance was higher, while transcript levels were slightly altered in roots exposed to CAN. This is the first report on identification of differentially nitrated proteins in response to supplementation with nonproteinogenic amino acid. Among nitrated proteins differentially modified by CAN, seed storage proteins (after short-term CAN treatment) and components of the cellular redox system (after prolonged CAN supplementation) were identified. The findings demonstrate that due to inhibition of NOS-like activity, CAN leads to modification in antioxidant system. Limitation in GSNO level is due to lower nitric oxide formation, while GSNO catabolism is less affected. We demonstrated that monodehydroascorbate reductase, activity of which is inhibited in roots of CAN-treated plants, is the protein preferentially modified by tyrosine nitration.

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Section: Methodsmentioning

confidence: 99%

Canavanine-Induced Decrease in Nitric Oxide Synthesis Alters Activity of Antioxidant System but Does Not Impact S-Nitrosoglutathione Catabolism in Tomato Roots

et al. 2019

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“…It could be, for example, S-nitrosoglutathione reductase (GSNOR), which according to the enzyme classification is part of the family of Zn-dependent class III alcohol dehydrogenases (EC 1.1.1.1), which are distinct from members of class I ADH family by their high affinity towards long-chain alcohols such as cinnamyl alcohol, farnesol, and geraniol. The main function of GSNOR is the reduction of S-nitrosoglutathione (GSNO) [41]. Increased expression of Pgb results in a decreased availability of free NO, and thus a reduced amount of GSNO.…”

Section: Energy-related Proteins and Metabolitesmentioning

confidence: 99%

The Hypoxic Proteome and Metabolome of Barley (Hordeum vulgare L.) with and without Phytoglobin Priming

Andrzejczak

Havelund

Wang

et al. 2020

IJMS

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Overexpression of phytoglobins (formerly plant hemoglobins) increases the survival rate of plant tissues under hypoxia stress by the following two known mechanisms: (1) scavenging of nitric oxide (NO) in the phytoglobin/NO cycle and (2) mimicking ethylene priming to hypoxia when NO scavenging activates transcription factors that are regulated by levels of NO and O2 in the N-end rule pathway. To map the cellular and metabolic effects of hypoxia in barley (Hordeum vulgare L., cv. Golden Promise), with or without priming to hypoxia, we studied the proteome and metabolome of wild type (WT) and hemoglobin overexpressing (HO) plants in normoxia and after 24 h hypoxia (WT24, HO24). The WT plants were more susceptible to hypoxia than HO plants. The chlorophyll a + b content was lowered by 50% and biomass by 30% in WT24 compared to WT, while HO plants were unaffected. We observed an increase in ROS production during hypoxia treatment in WT seedlings that was not observed in HO seedlings. We identified and quantified 9694 proteins out of which 1107 changed significantly in abundance. Many proteins, such as ion transporters, Ca2+-signal transduction, and proteins related to protein degradation were downregulated in HO plants during hypoxia, but not in WT plants. Changes in the levels of histones indicates that chromatin restructuring plays a role in the priming of hypoxia. We also identified and quantified 1470 metabolites, of which the abundance of >500 changed significantly. In summary the data confirm known mechanisms of hypoxia priming by ethylene priming and N-end rule activation; however, the data also indicate the existence of other mechanisms for hypoxia priming in plants.

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“…After centrifugation at 12,000× g and 4 °C for 15 min, supernatants were used for kinetic assays following Kubienová et al . (2016).…”

Section: Methodsmentioning

confidence: 99%

“…GSNOR extraction was performed on 200 mg fresh leaf disks ground in 1 ml 0.1 molÁl À1 phosphate buffer pH 7.2 with a mortar and pestle in an ice bath (adapted from Frungillo et al 2013). After centrifugation at 12,0009g and 4 °C for 15 min, supernatants were used for kinetic assays following Kubienov a et al (2016).…”

Section: Content Of Sno and S-nitrosoglutathione Reductase (Gsnor) Ac...mentioning

confidence: 99%

Drought memory in Acanthostachys strobilacea, a CAM epiphytic bromeliad

Carvalho,

Gaspar,

Nievola

2024

Plant Biol J

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Stress memory is the development of altered responses to stress due to previous exposure, which might result in increased tolerance. Biochemical and physiological parameters shown to be positively affected by stress memory include those of the antioxidant and nitrosative metabolism, photosynthetic pigments and osmolyte content. Epiphytic bromeliads likely present stress memory since they experience frequent droughts in the canopies. Thus, we aimed to evaluate if the epiphytic bromeliad Acanthostachys strobilacea (Schult. & Schult.f.) Klotzsch shows improved metabolic stress defence responses to a second drought and rewatering cycle compared to a single exposure. In a controlled environment chamber, 90‐day‐old plants were exposed to one or two drought–rewatering cycles of 14 days without irrigation and 5 days of rewatering each. Sampling occurred after the final drought and rewatering periods for one or two cycles treatments. The free amino acid, chlorophyll, and carotenoid levels and S‐nitrosoglutathione reductase (GSNOR) activity were higher at the second drought than at the first exposure. The rise in nocturnal acidification (indicative of increased CAM activity) caused by the initial drought persisted through the second drought–rewatering cycle, implying a lasting memory effect on CAM activity. Furthermore, the second recovery did not induce glutathione accumulation, as in the first rewatering event, suggesting the pre‐exposure to drought reduced this thiol's demand during a later recovery. Our results evidence metabolic changes related to drought stress memory in A. strobilacea, supporting this mechanism might be involved in the tolerance of epiphytic bromeliads to intermittent droughts.

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Detection of S-Nitrosoglutathione Reductase Activity in Plants

Cited by 4 publications

References 16 publications

Canavanine-Induced Decrease in Nitric Oxide Synthesis Alters Activity of Antioxidant System but Does Not Impact S-Nitrosoglutathione Catabolism in Tomato Roots

Canavanine-Induced Decrease in Nitric Oxide Synthesis Alters Activity of Antioxidant System but Does Not Impact S-Nitrosoglutathione Catabolism in Tomato Roots

The Hypoxic Proteome and Metabolome of Barley (Hordeum vulgare L.) with and without Phytoglobin Priming

Drought memory in Acanthostachys strobilacea, a CAM epiphytic bromeliad

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