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Literature summary extracted from

  • Kubienova, L.; Kopecny, D.; Tylichova, M.; Briozzo, P.; Skopalova, J.; Sebela, M.; Navratil, M.; Tache, R.; Luhova, L.; Barroso, J.B.; Petrivalsky, M.
    Structural and functional characterization of a plant S-nitrosoglutathione reductase from Solanum lycopersicum (2013), Biochimie, 95, 889-902.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.284 real-time PCR expression analysis, recombinant expression in Escherichia coli as N-terminally His-tagged enzyme Solanum lycopersicum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.284 purified recombinant His-tagged enzyme, in complex with NAD+ and with NADH, hanging drop vapour diffusion, mixing of 0.003 ml of 22 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 2.5 mM NAD+, or 2.5 mM NADH and 5.0 mM GSH, 20°C, X-ray diffraction tructure determination and analysis at 1.85-2.14 A resolution, molecular replacement and modeling Solanum lycopersicum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.284 2-mercaptoethanol inhibits both the reductase and dehydrogenase reactions by 30% at 1 mM Solanum lycopersicum
1.1.1.284 Ag+
-
Solanum lycopersicum
1.1.1.284 ascorbic acid inhibits both the reductase and dehydrogenase reactions by 30% at 1 mM Solanum lycopersicum
1.1.1.284 Decanoic acid
-
Solanum lycopersicum
1.1.1.284 dithiothreitol inhibits both the reductase and dehydrogenase reactions by 30% at 1 mM Solanum lycopersicum
1.1.1.284 dodecanoic acid noncompetitive Solanum lycopersicum
1.1.1.284 EDTA complete inactivation Solanum lycopersicum
1.1.1.284 glutathione noncompetitive inhibitor, inhibits GSNO reduction Solanum lycopersicum
1.1.1.284 Hg2+
-
Solanum lycopersicum
1.1.1.284 N6022 noncompetitive Solanum lycopersicum
1.1.1.284 octanoic acid
-
Solanum lycopersicum
1.1.1.284 S-Methylglutathione noncompetitive inhibitor, inhibits GSNO reduction Solanum lycopersicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.284 0.04
-
NAD+ pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum
1.1.1.284 0.057
-
S-nitrosylglutathione pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum
1.1.1.284 0.058
-
NADH pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum
1.1.1.284 0.058
-
S-(hydroxymethyl)glutathione pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.1.284 cytosol
-
Solanum lycopersicum 5829
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.284 additional information other monovalent and bivalent cations, besides Hg2+ or Ag+, have only a small effect Solanum lycopersicum
1.1.1.284 Zn2+ required for the oxidation reaction. The catalytic zinc ion functions as a Lewis acid, is bound by Cys177 and Cys47, His69 and either Glu70 or water (hydroxide anion). The structural zinc is bound by four cysteines, Cys99, Cys102, Cys105, and Cys113 Solanum lycopersicum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.284 40000
-
2 * 40000, SDS-PAGE Solanum lycopersicum
1.1.1.284 81090
-
native enzyme, sequence calculation Solanum lycopersicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.284 S-(hydroxymethyl)glutathione + NAD+ Solanum lycopersicum
-
S-formylglutathione + NADH + H+
-
?
1.1.1.284 S-(hydroxymethyl)glutathione + NAD+ Solanum lycopersicum Amateur
-
S-formylglutathione + NADH + H+
-
?
1.1.1.284 S-nitrosoglutathione + NADH Solanum lycopersicum
-
? + NAD+ a variety of products depending on cellular conditions, including glutathione disulfide, glutathione sulfinamide and hydroxylamine ?
1.1.1.284 S-nitrosoglutathione + NADH Solanum lycopersicum Amateur
-
? + NAD+ a variety of products depending on cellular conditions, including glutathione disulfide, glutathione sulfinamide and hydroxylamine ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.284 Solanum lycopersicum D2Y3F4
-
-
1.1.1.284 Solanum lycopersicum Amateur D2Y3F4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.284 recombinant N-terminally His-tagged enzyme from Escherichia coli by methal chelating chromatography Solanum lycopersicum

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.284 S-(hydroxymethyl)glutathione + NAD(P)+ = S-formylglutathione + NAD(P)H + H+ structure-function analysis, active site and ligand binding structures,reaction mechanism, overview Solanum lycopersicum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.284 cotyledon
-
Solanum lycopersicum
-
1.1.1.284 flower
-
Solanum lycopersicum
-
1.1.1.284 fruit the enzyme is highly expressed in the pistil and stamens and in fruits during ripening Solanum lycopersicum
-
1.1.1.284 leaf
-
Solanum lycopersicum
-
1.1.1.284 additional information expression is higher in roots and stems compared to leaves of young plants Solanum lycopersicum
-
1.1.1.284 pistil
-
Solanum lycopersicum
-
1.1.1.284 root
-
Solanum lycopersicum
-
1.1.1.284 shoot apex
-
Solanum lycopersicum
-
1.1.1.284 stamen
-
Solanum lycopersicum
-
1.1.1.284 stem
-
Solanum lycopersicum
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.284 additional information the reaction mechanism involved the oxidation of a hydroxyl group of S-(hydroxymethyl)glutathione, spontaneous adduct of formaldehyde and glutathione, to form S-formylglutathione. Substrate specificity with alcohols and omega-hydroxyfatty acids, overview Solanum lycopersicum ?
-
?
1.1.1.284 additional information the reaction mechanism involved the oxidation of a hydroxyl group of S-(hydroxymethyl)glutathione, spontaneous adduct of formaldehyde and glutathione, to form S-formylglutathione. Substrate specificity with alcohols and omega-hydroxyfatty acids, overview Solanum lycopersicum Amateur ?
-
?
1.1.1.284 S-(hydroxymethyl)glutathione + NAD+
-
Solanum lycopersicum S-formylglutathione + NADH + H+
-
?
1.1.1.284 S-(hydroxymethyl)glutathione + NAD+
-
Solanum lycopersicum Amateur S-formylglutathione + NADH + H+
-
?
1.1.1.284 S-nitrosoglutathione + NADH
-
Solanum lycopersicum ? + NAD+ a variety of products depending on cellular conditions, including glutathione disulfide, glutathione sulfinamide and hydroxylamine ?
1.1.1.284 S-nitrosoglutathione + NADH
-
Solanum lycopersicum Amateur ? + NAD+ a variety of products depending on cellular conditions, including glutathione disulfide, glutathione sulfinamide and hydroxylamine ?

Subunits

EC Number Subunits Comment Organism
1.1.1.284 homodimer 2 * 40000, SDS-PAGE Solanum lycopersicum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.284 GSNOR
-
Solanum lycopersicum
1.1.1.284 HMGSH dehydrogenase
-
Solanum lycopersicum
1.1.1.284 S-nitrosoglutathione reductase
-
Solanum lycopersicum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.284 30
-
assay at Solanum lycopersicum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.284 8
-
assay at Solanum lycopersicum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.284 0.00043
-
N6022 pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum
1.1.1.284 0.76
-
dodecanoic acid pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.1.1.284 0.0003
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum N6022
1.1.1.284 0.526
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum dodecanoic acid
1.1.1.284 3.9
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum glutathione
1.1.1.284 5.1
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum Decanoic acid
1.1.1.284 5.8
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum S-Methylglutathione
1.1.1.284 6.1
-
pH 8.0, 30°C, recombinant enzyme Solanum lycopersicum octanoic acid

General Information

EC Number General Information Comment Organism
1.1.1.284 evolution the enzyme belongs to the large alcohol dehydrogenase superfamily, namely to the class III ADHs Solanum lycopersicum
1.1.1.284 additional information structure-function analysis, overview Solanum lycopersicum
1.1.1.284 physiological function in plants, GSNOR plays an important role in biotic and abiotic stress responses. S-nitrosylglutathione serves as a nitric oxide reservoir locally or possibly as NO donor in distant cells and tissues. NO and NO-related molecules such as S-nitrosothiols play a central role in the regulation of normal plant physiological processes and host defence. The key enzyme participates in the cellular homeostasis of S-NOs and in the metabolism of reactive nitrogen species Solanum lycopersicum