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Literature summary for 1.14.13.39 extracted from

  • Seckler, J.M.; Shen, J.; Lewis, T.H.J.; Abdulameer, M.A.; Zaman, K.; Palmer, L.A.; Bates, J.N.; Jenkins, M.W.; Lewis, S.J.
    NADPH diaphorase detects S-nitrosylated proteins in aldehyde-treated biological tissues (2020), Sci. Rep., 10, 21088 .
    View publication on PubMedView publication on EuropePMC

Inhibitors

Inhibitors Comment Organism Structure
additional information paraformaldehyde fixation of rat brain abolishes NOS activity in particulate and cytosolic fractions Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ in heme Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 L-arginine + 3 NADPH + 3 H+ + 4 O2 Rattus norvegicus overall reaction 2 L-citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
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?
2 L-arginine + 3 NADPH + 3 H+ + 4 O2 Rattus norvegicus Sprague-Dawley overall reaction 2 L-citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
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?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P29476
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Rattus norvegicus Sprague-Dawley P29476
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Source Tissue

Source Tissue Comment Organism Textmining
brain
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Rattus norvegicus
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brain stem
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Rattus norvegicus
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cerebellum
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Rattus norvegicus
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ganglion coeliac ganglia Rattus norvegicus
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additional information determination of NADPH diaphorase staining in preganglionic terminals, staining generally ascribed to NADH diaphorase activity in aldehyde-treated tissues Rattus norvegicus
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 L-arginine + 3 NADPH + 3 H+ + 4 O2 overall reaction Rattus norvegicus 2 L-citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
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?
2 L-arginine + 3 NADPH + 3 H+ + 4 O2 overall reaction Rattus norvegicus Sprague-Dawley 2 L-citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
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?
additional information the similar spatial distribution of NADPH diaphorase and nitric oxide synthase (NOS) in aldehyde-treated tissues has lead to the postulation that the catalytic activity of NOS promotes NADPH-dependent reduction of nitro-blue tetrazolium (NBT) to insoluble readily visualized diformazan particles. Since NADPH cannot directly reduce NBT, it is argued that NBT binds to NOS at the flavin electron transport domain and is reduced by a NOS form, which is previously reduced by beta-NADPH. However, paraformaldehyde fixation of rat brain abolishes NOS activity in particulate and cytosolic fractions and although fixation abolishes NADPH diaphorase in the particulate fraction, 50-60% of NADPH diaphorase activity remains in the cytosolic fraction. This suggests that cytosolic NADPH diaphorase in aldehyde-treated tissues results from a factor other than NOS. NOS contains heme iron, flavin adenine dinucleotide and flavin mononucleotide. NADPH-dependent reduction of NBT in aldehyde-treated tissues is not directly due to NOS because metal chelators that inhibit activity of metalloenzymes and deflavinating agents do not eliminate NADPH diaphorase. While the NOS active site is not required to promote the NADPH-dependent reduction of NBT, some other reactive site of the protein could still be involved. SNOs exist in cytoplasmic vesicles of endothelial cells of rat small mesenteric arteries, SNOs promote the NADPH-dependent reduction of NBT, both alpha-NADPH and beta-NADPH promote the reduction of NBT to diformazan and since alpha-NADPH will not donate electrons to NOS, it is unlikely that diformazan is the result of an increase in the catalytic activity of NOS, prior depletion of SNOs in tissues markedly reduces subsequent NADPH diaphorase staining. NADPH diaphorase activity in aldehyde-treated tissues is due to preformed pools of SNOs that facilitate NADPH-dependent reduction of NBT to diformazan Rattus norvegicus ?
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additional information the similar spatial distribution of NADPH diaphorase and nitric oxide synthase (NOS) in aldehyde-treated tissues has lead to the postulation that the catalytic activity of NOS promotes NADPH-dependent reduction of nitro-blue tetrazolium (NBT) to insoluble readily visualized diformazan particles. Since NADPH cannot directly reduce NBT, it is argued that NBT binds to NOS at the flavin electron transport domain and is reduced by a NOS form, which is previously reduced by beta-NADPH. However, paraformaldehyde fixation of rat brain abolishes NOS activity in particulate and cytosolic fractions and although fixation abolishes NADPH diaphorase in the particulate fraction, 50-60% of NADPH diaphorase activity remains in the cytosolic fraction. This suggests that cytosolic NADPH diaphorase in aldehyde-treated tissues results from a factor other than NOS. NOS contains heme iron, flavin adenine dinucleotide and flavin mononucleotide. NADPH-dependent reduction of NBT in aldehyde-treated tissues is not directly due to NOS because metal chelators that inhibit activity of metalloenzymes and deflavinating agents do not eliminate NADPH diaphorase. While the NOS active site is not required to promote the NADPH-dependent reduction of NBT, some other reactive site of the protein could still be involved. SNOs exist in cytoplasmic vesicles of endothelial cells of rat small mesenteric arteries, SNOs promote the NADPH-dependent reduction of NBT, both alpha-NADPH and beta-NADPH promote the reduction of NBT to diformazan and since alpha-NADPH will not donate electrons to NOS, it is unlikely that diformazan is the result of an increase in the catalytic activity of NOS, prior depletion of SNOs in tissues markedly reduces subsequent NADPH diaphorase staining. NADPH diaphorase activity in aldehyde-treated tissues is due to preformed pools of SNOs that facilitate NADPH-dependent reduction of NBT to diformazan Rattus norvegicus Sprague-Dawley ?
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-

Synonyms

Synonyms Comment Organism
NADPH diaphorase
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Rattus norvegicus
NADPH-d
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Rattus norvegicus
nNOS
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Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
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assay at room temperature Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
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assay at Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
beta-NADPH
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Rattus norvegicus
FAD
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Rattus norvegicus
heme
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Rattus norvegicus