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Information on EC 1.14.13.39 - nitric-oxide synthase (NADPH)

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IUBMB Comments
The enzyme consists of linked oxygenase and reductase domains. The eukaryotic enzyme binds FAD, FMN, heme (iron protoporphyrin IX) and tetrahydrobiopterin, and its two domains are linked via a regulatory calmodulin-binding domain. Upon calcium-induced calmodulin binding, the reductase and oxygenase domains form a complex, allowing electrons to flow from NADPH via FAD and FMN to the active center. The reductase domain of the enzyme from the bacterium Sorangium cellulosum utilizes a [2Fe-2S] cluster to transfer the electrons from NADPH to the active center. cf. EC 1.14.14.47, nitric-oxide synthase (flavodoxin).
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UNIPROT: P29475
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Word Map
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
hide(Overall reactions are displayed. Show all >>)
Synonyms
nos, inducible nitric oxide synthase, endothelial nitric oxide synthase, inducible no synthase, neuronal nitric oxide synthase, inducible nos, endothelial no synthase, endothelial nos, neuronal nos, no-synthase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
neuronal nitric oxide synthase
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neuronal NO synthase
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neuronal NOS
isoform
nitric oxide synthase
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NO synthase type I
nNOS
endothelium-derived relaxation factor-forming enzyme
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endothelium-derived relaxing factor synthase
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NADPH-diaphorase
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nitric oxide synthase
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nitric oxide synthetase
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NO synthase
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synthetase, nitric oxide
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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-
-
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oxidation
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-
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reduction
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SYSTEMATIC NAME
IUBMB Comments
L-arginine,NADPH:oxygen oxidoreductase (nitric-oxide-forming)
The enzyme consists of linked oxygenase and reductase domains. The eukaryotic enzyme binds FAD, FMN, heme (iron protoporphyrin IX) and tetrahydrobiopterin, and its two domains are linked via a regulatory calmodulin-binding domain. Upon calcium-induced calmodulin binding, the reductase and oxygenase domains form a complex, allowing electrons to flow from NADPH via FAD and FMN to the active center. The reductase domain of the enzyme from the bacterium Sorangium cellulosum utilizes a [2Fe-2S] cluster to transfer the electrons from NADPH to the active center. cf. EC 1.14.14.47, nitric-oxide synthase (flavodoxin).
CAS REGISTRY NUMBER
COMMENTARY hide
125978-95-2
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 L-arginine + 3 NADPH + 3 H+ + 4 O2
2 citrulline + 2 nitric oxide + 3 NADP+ + 4 H2O
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(6R)-5,6,7,8-tetrahydro-L-biopterin
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NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
the raise of intracellular calcium levels increases nNOS dephosphorylation and enzymatic activity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4S)-N-(4-amino-5-[aminoethyl]aminopentyl)-N''-nitroguanidine
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3-bromo-7-nitroindazole
nNOS-specific inhibitor, complete inhibition at 0.01 mM
3-[cis-4'-[(6''-aminopyridin-2''-yl)methyl]pyrrolidin-3'-ylamino]propan-1-ol
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L-N-methylarginine
NOS inhibitor, complete inhibition at 0.5 mM
L-Nomega-nitroarginine-(4R)-amino-L-proline amide
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L-Nomega-nitroarginine-2,4-L-diaminobutyramide
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N1-[cis-4'-[(6''-amino-4''-methylpyridin-2''-yl)methyl]pyrrolidin-3'-yl]-N2-(4'-chlorobenzyl)ethane-1,2-diamine
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N1-[cis-4'-[(6''-aminopyridin-2''-yl)methyl]pyrrolidin-3'-yl]ethane-1,2-diamine
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N1-[trans-4'-[(6''-amino-4''-methylpyridin-2''-yl)methyl]pyrrolidin-3'-yl]-N2-(3'-chlorobenzyl)ethane-1,2-diamine
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0019
L-arginine
mutant nNOS D597N/M336V
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00015
(4S)-N-(4-amino-5-[aminoethyl]aminopentyl)-N''-nitroguanidine
wild type nNOS
0.0094 - 0.0492
3-[cis-4'-[(6''-aminopyridin-2''-yl)methyl]pyrrolidin-3'-ylamino]propan-1-ol
0.0001
L-Nomega-nitroarginine-(4R)-amino-L-proline amide
wild type nNOS
0.0003
L-Nomega-nitroarginine-2,4-L-diaminobutyramide
wild type nNOS
0.000085 - 0.0012
N1-[cis-4'-[(6''-amino-4''-methylpyridin-2''-yl)methyl]pyrrolidin-3'-yl]-N2-(4'-chlorobenzyl)ethane-1,2-diamine
0.000388 - 0.0367
N1-[cis-4'-[(6''-aminopyridin-2''-yl)methyl]pyrrolidin-3'-yl]ethane-1,2-diamine
0.00025 - 0.0061
N1-[trans-4'-[(6''-amino-4''-methylpyridin-2''-yl)methyl]pyrrolidin-3'-yl]-N2-(3'-chlorobenzyl)ethane-1,2-diamine
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
nNOS is more frequently expressed in normal than in osteoarthritic chondrocytes
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
NOS1_HUMAN
1434
0
160970
Swiss-Prot
other Location (Reliability: 3)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15000
x * 15000, SDS-PAGE
150000
SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 15000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D597N/M336V
mutant of nNOS, the Ki values for the nNOS double mutant increase for all inhibitors but the mutant still binds these inhibitors better than eNOS
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni Sepharose column chromatography, 2',5'-ADP Sepharose column chromatography, and Superdex 200 gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in SH-SY5Y cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
lung tissue from patients with severe and very severe chronic obstructive pulmonary disease have graded increases in nNOS (mRNA and protein) compared with non-smokers and normal smokers
neither interleukin-1 nor tissue necrosis factor-alpha is capable of inducing nNOS synthesis for up to 48 h treatment, treatment with insulin-like growth factor-1 or transforming growth factor-beta for up to 48 h has no effect on the levels of nNOS
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Brindicci, C.; Kharitonov, S.A.; Ito, M.; Elliott, M.W.; Hogg, J.C.; Barnes, P.J.; Ito, K.
Nitric oxide synthase isoenzyme expression and activity in peripheral lungs of COPD patients
Am. J. Respir. Crit. Care Med.
181
21-30
2009
Homo sapiens (P29474), Homo sapiens (P29475), Homo sapiens (P35228), Homo sapiens
Manually annotated by BRENDA team
Mikula, I.; Durocher, S.; Martasek, P.; Mutus, B.; Slama-Schwok, A.
Isoform-specific differences in the nitrite reductase activity of nitric oxide synthases under hypoxia
Biochem. J.
418
673-682
2009
Homo sapiens, Homo sapiens (P29474), Homo sapiens (P29475)
Manually annotated by BRENDA team
Igarashi, J.; Li, H.; Jamal, J.; Ji, H.; Fang, J.; Lawton, G.R.; Silverman, R.B.; Poulos, T.L.
Crystal structures of constitutive nitric oxide synthases in complex with de novo designed inhibitors
J. Med. Chem.
52
2060-2066
2009
Homo sapiens (P29474), Homo sapiens (P29475)
Manually annotated by BRENDA team
Aquilano, K.; Baldelli, S.; Rotilio, G.; Ciriolo, M.R.
Role of nitric oxide synthases in Parkinsons disease: a review on the antioxidant and anti-inflammatory activity of polyphenols
Neurochem. Res.
33
2416-2426
2008
Homo sapiens, Homo sapiens (P29475), Homo sapiens (P35228)
Manually annotated by BRENDA team
Rosa, S.C.; Judas, F.; Lopes, M.C.; Mendes, A.F.
Nitric oxide synthase isoforms and NF-kappaB activity in normal and osteoarthritic human chondrocytes: regulation by inducible nitric oxide
Nitric Oxide
19
276-283
2008
Homo sapiens (P29475), Homo sapiens (P35228), Homo sapiens
Manually annotated by BRENDA team