NOS3

Summary

NOS3 (nitric oxide synthase 3, HGNC:7876) is a protein-coding gene on chromosome 7q36.1, encoding Nitric oxide synthase 3 (P29474). Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway.

Nitric oxide is a reactive free radical which acts as a biologic mediator in several processes, including neurotransmission and antimicrobial and antitumoral activities. Nitric oxide is synthesized from L-arginine by nitric oxide synthases. Variations in this gene are associated with susceptibility to coronary spasm. Alternative splicing and the use of alternative promoters results in multiple transcript variants.

Source: NCBI Gene 4846 — RefSeq curated summary.

At a glance

• Clinical variants (ClinVar): 191 total — 2 pathogenic, 2 likely-pathogenic
• GTEx eGene: eQTL in 16 tissues, sQTL in 14 (q < 0.05)
• Drug target: yes — 1 via a complex or family; 6 molecules with ChEMBL bioactivity
• Constraint (gnomAD): pLI 0.00 (LoF-tolerant), LOEUF 0.90 (unconstrained)
• MANE Select transcript: NM_000603
• Interaction hub: 2,920 STRING partners (top 3% of genes corpus-wide)
• Notable: 191 clinical variants but no curated precision-oncology (CIViC) evidence yet

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:7876
Approved symbol	NOS3
Name	nitric oxide synthase 3
Location	7q36.1
Locus type	gene with protein product
Status	Approved
Aliases	ECNOS, eNOS
Ensembl gene	ENSG00000164867
Ensembl biotype	protein_coding
OMIM	163729
Entrez	4846

Gene structure

Transcript identifiers

Ensembl transcripts: 18 — 11 protein_coding, 4 protein_coding_CDS_not_defined, 2 nonsense_mediated_decay, 1 retained_intron

MANE Select: ENST00000297494

Transcript support level (Ensembl TSL; 1 = fully mRNA-supported): TSL 1: 3, TSL 2: 4, TSL 3: 2, TSL 5: 1

ENST00000297494, ENST00000460603, ENST00000461406, ENST00000467517, ENST00000468293, ENST00000473057, ENST00000475017, ENST00000475454, ENST00000477227, ENST00000484524, ENST00000908206, ENST00000943232, ENST00000943234, ENST00001009542, ENST00001009543, ENST00001109056, ENST00001126102, ENST00001135338

RefSeq mRNA: 4 — MANE Select: NM_000603 NM_000603, NM_001160109, NM_001160110, NM_001160111

CCDS: CCDS55182, CCDS55183, CCDS5912

Canonical transcript exons

ENST00000297494 — 27 exons

Exon	Start	End
ENSE00001088117	151009398	151009585
ENSE00001088120	151006427	151006494
ENSE00001088124	151006889	151007005
ENSE00001088128	151007102	151007276
ENSE00001088131	150998539	150998680
ENSE00001088133	151001231	151001425
ENSE00001088137	151010597	151010807
ENSE00001088139	150998946	150999085
ENSE00001088143	151001821	151001965
ENSE00001088144	151010115	151010287
ENSE00001088150	150999190	150999364
ENSE00001088154	151000498	151000599
ENSE00001088158	150996763	150996925
ENSE00001088160	150996404	150996552
ENSE00001088161	151008930	151009062
ENSE00001088163	151001544	151001617
ENSE00001141355	151014008	151014588
ENSE00001141361	150993753	150993961
ENSE00001851539	150991017	150991300
ENSE00003463030	151013724	151013918
ENSE00003463776	151013231	151013379
ENSE00003545944	150995203	150995314
ENSE00003585672	151010899	151010986
ENSE00003622733	151012351	151012472
ENSE00003634102	151009189	151009267
ENSE00003642116	150998357	150998448
ENSE00003648730	151002200	151002304

Expression profiles

Bgee: expression breadth ubiquitous, 168 present calls, max score 96.16.

FANTOM5 (CAGE): breadth broad, TPM avg 15.3641 / max 479.1941, expressed in 887 samples.

Highest expression (Bgee): spleen, apex of heart, lower esophagus mucosa, omental fat pad, peritoneum.

Genetic regulation across tissues (GTEx eQTL / sQTL)

NOS3 is an eGene in GTEx v10 — its expression is genetically regulated (eQTL) in 16 tissues and its splicing (sQTL) in 14 (q < 0.05). Strongest associations first; slope = normalized effect of the lead variant’s alternative allele.

showing 20 of 30 significant tissue × QTL associations:

Tissue	QTL	q-value	Slope	Lead variant
Testis	sqtl	6.6e-38	+1.42	7:151000206:A:G
Adipose Visceral Omentum	sqtl	4.5e-26	-1.01	7:151000206:A:G
Spleen	sqtl	3.5e-18	-1.21	7:151000206:A:G
Thyroid	sqtl	1.1e-16	-0.81	7:151000206:A:G
Heart Left Ventricle	sqtl	8.9e-13	-0.96	7:151000206:A:G
Nerve Tibial	sqtl	2.5e-12	-0.75	7:151000206:A:G
Cells Cultured fibroblasts	eqtl	3.9e-12	-0.40	7:151009295:C:A
Heart Atrial Appendage	sqtl	5.7e-12	-0.88	7:151000206:A:G
Cells Cultured fibroblasts	sqtl	2.3e-07	+0.44	7:151009827:G:A
Lung	sqtl	1.4e-06	-0.55	7:151000206:A:G
Prostate	sqtl	8.0e-05	-0.86	7:151000206:A:G
Whole Blood	eqtl	2.2e-04	-0.24	7:150995356:G:A
Esophagus Muscularis	sqtl	3.3e-04	-0.52	7:151000206:A:G
Skin Not Sun Exposed Suprapubic	eqtl	5.3e-04	+0.17	7:150970982:C:T
Esophagus Mucosa	eqtl	6.9e-04	+0.15	7:151090363:T:G
Heart Atrial Appendage	eqtl	1.1e-03	-0.31	7:150993088:C:T
Esophagus Muscularis	eqtl	1.3e-03	-0.29	7:150993088:C:T
Lung	eqtl	1.9e-03	-0.35	7:150993088:C:T
Adipose Visceral Omentum	eqtl	2.0e-03	-0.28	7:150993088:C:T
Skin Sun Exposed Lower leg	eqtl	2.6e-03	-0.28	7:150993088:C:T

FANTOM5 promoters

showing 10 of 12 alternative TSS:

Promoter ID	TPM avg	Samples expressed
82024	9.6081	412
82027	1.4593	304
82028	1.3921	251
82023	1.0436	320
82030	0.8136	179
82031	0.7703	135
82019	0.0849	23
82022	0.0770	44
82029	0.0425	18
82020	0.0323	10

Top tissues by expression

showing 100 of 294 tissues by Bgee expression score (0-100, higher = more expressed):

Tissue	Anatomy ID	Expression score	Quality
spleen	UBERON:0002106	96.16	gold quality
apex of heart	UBERON:0002098	93.22	gold quality
lower esophagus mucosa	UBERON:0035834	90.66	gold quality
omental fat pad	UBERON:0010414	90.15	gold quality
peritoneum	UBERON:0002358	90.02	gold quality
right atrium auricular region	UBERON:0006631	89.55	gold quality
metanephros cortex	UBERON:0010533	88.23	gold quality
adipose tissue of abdominal region	UBERON:0007808	87.69	gold quality
upper lobe of left lung	UBERON:0008952	87.28	gold quality
left uterine tube	UBERON:0001303	87.12	gold quality
heart left ventricle	UBERON:0002084	86.77	gold quality
body of uterus	UBERON:0009853	86.58	gold quality
cardiac atrium	UBERON:0002081	86.12	gold quality
right lobe of thyroid gland	UBERON:0001119	85.93	gold quality
left coronary artery	UBERON:0001626	85.85	gold quality
right coronary artery	UBERON:0001625	85.84	gold quality
cardiac ventricle	UBERON:0002082	85.67	gold quality
mucosa of stomach	UBERON:0001199	85.65	gold quality
tibial nerve	UBERON:0001323	85.45	gold quality
right lung	UBERON:0002167	85.02	gold quality
left lobe of thyroid gland	UBERON:0001120	84.47	gold quality
heart	UBERON:0000948	83.88	gold quality
upper lobe of lung	UBERON:0008948	83.66	gold quality
primordial germ cell in gonad	CL:0000670 ∩ UBERON:0000991	83.51	gold quality
esophagogastric junction muscularis propria	UBERON:0035841	83.41	gold quality
coronary artery	UBERON:0001621	83.39	gold quality
adenohypophysis	UBERON:0002196	83.38	gold quality
left testis	UBERON:0004533	83.33	gold quality
ectocervix	UBERON:0012249	83.21	gold quality
sural nerve	UBERON:0015488	83.18	gold quality
C1 segment of cervical spinal cord	UBERON:0006469	83.05	gold quality
right testis	UBERON:0004534	82.99	gold quality
lower esophagus muscularis layer	UBERON:0035833	82.95	gold quality
lower esophagus	UBERON:0013473	82.93	gold quality
hindlimb stylopod muscle	UBERON:0004252	82.80	gold quality
endocervix	UBERON:0000458	82.74	gold quality
thyroid gland	UBERON:0002046	82.44	gold quality
subcutaneous adipose tissue	UBERON:0002190	81.61	gold quality
gall bladder	UBERON:0002110	81.57	gold quality
left adrenal gland cortex	UBERON:0035825	81.44	gold quality
right adrenal gland	UBERON:0001233	80.86	gold quality
right frontal lobe	UBERON:0002810	80.68	gold quality
left adrenal gland	UBERON:0001234	80.65	gold quality
body of stomach	UBERON:0001161	80.50	gold quality
right lobe of liver	UBERON:0001114	80.40	gold quality
muscle layer of sigmoid colon	UBERON:0035805	79.94	gold quality
right adrenal gland cortex	UBERON:0035827	79.83	gold quality
testis	UBERON:0000473	79.79	gold quality
small intestine Peyer’s patch	UBERON:0003454	79.78	gold quality
esophagus	UBERON:0001043	79.71	gold quality
ascending aorta	UBERON:0001496	79.35	gold quality
thoracic aorta	UBERON:0001515	79.08	gold quality
transverse colon	UBERON:0001157	78.65	gold quality
pituitary gland	UBERON:0000007	78.63	gold quality
spinal cord	UBERON:0002240	78.62	gold quality
vagina	UBERON:0000996	78.58	gold quality
aorta	UBERON:0000947	78.52	gold quality
tibial artery	UBERON:0007610	78.50	gold quality
popliteal artery	UBERON:0002250	78.45	gold quality
right ovary	UBERON:0002118	78.36	gold quality
gastrocnemius	UBERON:0001388	78.31	gold quality
adrenal cortex	UBERON:0001235	78.27	gold quality
minor salivary gland	UBERON:0001830	78.06	gold quality
adrenal gland	UBERON:0002369	77.77	gold quality
mucosa of transverse colon	UBERON:0004991	77.73	gold quality
esophagus mucosa	UBERON:0002469	77.63	gold quality
male germ line stem cell (sensu Vertebrata) in testis	CL:0000089 ∩ UBERON:0000473	77.47	gold quality
muscle of leg	UBERON:0001383	77.25	gold quality
stomach	UBERON:0000945	76.86	gold quality
skin of abdomen	UBERON:0001416	76.79	gold quality
cingulate cortex	UBERON:0003027	76.78	gold quality
skin of leg	UBERON:0001511	76.63	gold quality
anterior cingulate cortex	UBERON:0009835	76.51	gold quality
colonic epithelium	UBERON:0000397	76.19	gold quality
amygdala	UBERON:0001876	76.17	gold quality
cortical plate	UBERON:0005343	76.04	gold quality
descending thoracic aorta	UBERON:0002345	75.86	gold quality
small intestine	UBERON:0002108	75.80	gold quality
putamen	UBERON:0001874	75.65	gold quality
adipose tissue	UBERON:0001013	75.27	gold quality
left ovary	UBERON:0002119	75.24	gold quality
caudate nucleus	UBERON:0001873	74.39	gold quality
prefrontal cortex	UBERON:0000451	74.33	gold quality
connective tissue	UBERON:0002384	74.08	gold quality
olfactory segment of nasal mucosa	UBERON:0005386	74.01	gold quality
hypothalamus	UBERON:0001898	73.92	gold quality
prostate gland	UBERON:0002367	73.92	gold quality
fundus of stomach	UBERON:0001160	73.80	gold quality
adult mammalian kidney	UBERON:0000082	73.47	gold quality
nucleus accumbens	UBERON:0001882	73.34	gold quality
granulocyte	CL:0000094	73.30	gold quality
vermiform appendix	UBERON:0001154	73.24	gold quality
mouth mucosa	UBERON:0003729	73.12	gold quality
body of pancreas	UBERON:0001150	72.90	gold quality
colon	UBERON:0001155	72.64	gold quality
sigmoid colon	UBERON:0001159	72.61	gold quality
right hemisphere of cerebellum	UBERON:0014890	72.60	gold quality
saliva-secreting gland	UBERON:0001044	72.53	gold quality
right uterine tube	UBERON:0001302	72.42	gold quality
intestine	UBERON:0000160	72.24	gold quality

Single-cell (SCXA)

Detected in 1 experiment(s), a significant marker in 1.

Experiment	Marker?	Max mean expression
E-ANND-3	yes	8.01

HPA expression

RNA tissue specificity: Tissue enhanced

showing 60 of 119 HPA tissues / cells by nTPM:

Tissue / cell	Axis	nTPM	Protein (IHC)
Spleen	tissue	49.4	not detected
Locus coeruleus	tissue	12.6
Substantia nigra	tissue	12.4
Superior colliculi	tissue	11.9
Hippocampus	tissue	11.4	not detected
Thalamus	tissue	11.2
Nucleus accumbens	tissue	9.6
Pons	tissue	9.3
Midbrain	tissue	9.2
Cerebral cortex	tissue	8.4	low
Adipose tissue	tissue	8.4	not detected
Choroid plexus	tissue	8.4
Insular cortex	tissue	8.4
Medulla oblongata	tissue	8.3
Somatosensory cortex	tissue	8.2
Postcentral gyrus	tissue	8.1
Spinal cord	tissue	8.0
Motor cortex	tissue	7.8
Hippocampal formation	tissue	7.8
Entorhinal cortex	tissue	7.8
Prefrontal cortex	tissue	7.6
Retrosplenial and cingulate cortex	tissue	7.6
Putamen	tissue	7.4
Temporal cortex	tissue	7.4
Pituitary gland	tissue	7.0
Vagina	tissue	7.0	not detected
Retrosplenial cortex	tissue	6.8
Occipital cortex	tissue	6.7
Basal ganglia	tissue	6.6	not detected
Septum	tissue	6.6
Ventral forebrain	tissue	6.5
Corpus callosum	tissue	6.1
Heart muscle	tissue	6.0	not detected
Hypothalamus	tissue	5.8
Amygdala	tissue	5.6
Periaqueductal gray	tissue	5.5
Appendix	tissue	5.5	not detected
Postcentral gyrus	tissue	4.9
Olfactory bulb	tissue	4.6
Frontal cortex	tissue	4.5
Placenta	tissue	4.5	high
Gallbladder	tissue	4.4	not detected
Urinary bladder	tissue	3.7	not detected
Cerebellum	tissue	3.7	not detected
Endometrium 1	tissue	3.6	not detected
Smooth muscle	tissue	3.5	not detected
Caudate putamen	tissue	3.5
Tongue	tissue	3.3
Lung	tissue	3.3	not detected
Skin 1	tissue	3.2	not detected
Esophagus	tissue	3.0	not detected
Skeletal muscle	tissue	2.9	not detected
Cerebellum	tissue	2.9	not detected
Testis	tissue	2.8	not detected
Adrenal gland	tissue	2.7	not detected
Duodenum	tissue	2.5	not detected
Lymph node	tissue	2.4	not detected
Pancreas	tissue	2.4	not detected
Cervix	tissue	2.3	not detected
Testis	tissue	2.3	medium

Brain sub-regions (HPA Brain Atlas): detected in 377; highest nTPM — Posterior thalamic nucleus (25.0), Nucleus cuneatus (19.9), Dorsal cochlear nucleus (19.6), Reticulotegmental nucleus (19.1), Lateral amygdala (19.0).

Regulation

Is transcription factor: no

miRNA regulators (miRDB)

Targeting NOS3 by miRDB confidence (max_score; target_count = how many genes the miRNA targets in total — lower means more specific).

14 miRNAs:

miRNA	Max score	Avg score	miRNA target_count
HSA-MIR-6758-5P	100.00	66.21	1470
HSA-MIR-6856-5P	100.00	65.47	1298
HSA-MIR-4650-5P	99.98	64.69	999
HSA-MIR-3202	99.66	67.70	2737
HSA-MIR-1303	99.65	69.77	1662
HSA-MIR-766-5P	99.47	67.91	2225
HSA-MIR-4254	99.11	65.15	1315
HSA-MIR-4324	99.04	70.14	1569
HSA-MIR-4297	98.77	66.95	2013
HSA-MIR-6873-5P	98.45	66.14	1417
HSA-MIR-7156-3P	98.25	67.66	859
HSA-MIR-5581-5P	97.91	66.50	965
HSA-MIR-4638-3P	97.90	65.75	905
HSA-MIR-4448	97.04	66.22	752

Functional genomics

gnomAD constraint: pLI 2.55e-25, LOEUF 0.9, missense Z 1.49 — population loss-of-function intolerance (pLI ≥ 0.9 = LoF-intolerant; lower LOEUF = more constrained). gnomAD

Regional missense constraint (gnomAD)

ENST00000297494 splits into 5 regions with different missense tolerance; 0 are missense-constrained. Missense variants in a constrained region (low observed/expected) are more likely to be pathogenic. gnomAD

Region (GRCh38)	Missense obs / exp	o/e	Constrained
7:150,993,804–150,998,678	374 / 377.0	0.99	no
7:150,998,679–150,999,009	12 / 30.1	0.40	no
7:150,999,010–151,010,672	745 / 853.9	0.87	no
7:151,010,673–151,013,274	106 / 167.7	0.63	no
7:151,013,275–151,014,166	228 / 227.7	1.00	no

Literature-anchored findings (GeneRIF, showing 40)

• data fall short of showing significant association between a variant of the promoter of interleukin-1beta, polymorphism of angiotensinogen, and the missense variant of endothelial nitric oxide synthase and occurrence of idiopathic recurrent miscarriage (PMID:11756575)
• Glu298Asp polymorphism is associated with a 5 year lower mean age at end stage renal disease in males with AD polycytic kidneys. (PMID:11823442)
• Endothelial nitric oxide gene polymorphism (Glu298Asp) is not associated with coronary artery disease in Turkish population. (PMID:11858501)
• The human alveolar type II epithelium-like cell line A549 expresses nitric oxide synthase type 2 (NOS2), but not NOS3. (PMID:11880293)
• Effect of ecNOS polymorphisms and coronary artery disease upon exhaled nitric oxide (PMID:11894144)
• Association of a polymorphism of the ecNOS gene with myocardial infarction in a subgroup of Turkish MI patients (PMID:11903359)
• Play a key role in regulation of airway function and in the pathophysiology of inflammation in IgE-mediated allergic disease. (PMID:11906374)
• Endothelial nitric oxide synthase gene polymorphisms in Behcet’s disease. (PMID:11908569)
• Polymorphism of the endothelial nitric oxide synthase predicts risk for severe diabetic retinopathy. (enos4) (PMID:11918626)
• the role and intracellular signal pathway of endothelial nitric oxide synthase (eNOS) activation induced by VEGF (PMID:11961297)
• Increased risk of systemic sclerosis in eNOS 894T allele carriers. (PMID:12015245)
• Data provides evidence for association between preeclampsia and a polymorphism of the NOS3 gene. (PMID:12044319)
• Alkalosis stimulates endothelial nitric oxide synthase in cultured human pulmonary arterial endothelial cells. (PMID:12060567)
• Glu298Asp polymorphism is associated with differences in endothelial responses to both smoking and n-3 fatty acids in healthy young subjects, and these findings raise the possibility of genotype-specific prevention strategies in cardiovascular disease. (PMID:12065317)
• the a allele of the ecNOS gene polymorphism showed a significantly higher incidence among patients with end-stage renal failure caused by autosomal dominant polycystic kidney disease (PMID:12077489)
• in addition to alpha-gal A gene mutations, eNOS genetic variations are implicated in Fabry’s disease (PMID:12121349)
• ecNOS gene polymorphism is associated with endothelium-dependent vasodilation in Type 2 diabetes. (PMID:12124201)
• effects of gene polymorphisms on platelet function, nitric oxide release, and interactions with estradiol (PMID:12142730)
• Polymorphisms of the eNOS gene were significantly associated with high-altitude pulmonary edema (PMID:12176955)
• association of not-GG genotype with a-allele from ecNOS4a/b polymorphism is related to significant increase in susceptibility to prostate cancer (PMID:12195160)
• Smoking status-dependent association of the 27-bp repeat polymorphism in intron 4 of endothelial nitric oxide synthase gene with plasma nitric oxide concentrations. (PMID:12204432)
• This gene 4a/b polymorphism was not associated with the extent of coronary arteriosclerosis, but the a-allele of the variant seems to protect to some degree against the development of myocardial infarction. (PMID:12226742)
• human enos binds to porin, a voltage-dependent anion channel (PMID:12228731)
• C-reactive protein decreases eNOS expression and bioactivity in human aortic endothelial cells (PMID:12234944)
• The ecNOS Glu298Asp polymorphism is associated with reduced basal NO production and might therefore have functional implications in the development of atherosclerosis or hypertension. (PMID:12359981)
• assessment of association of human endothelial nitric oxide synthase gene polymorphism with the myocardial infarction (PMID:12362496)
• conclude that an increase in eNOS activity is beneficial and provides protection against atherosclerosis (PMID:12364322)
• Association of eNOS Glu298Asp polymorphism with end-stage renal disease (PMID:12364359)
• in patients with endometriosis a significant negative correlation was observed between luminal expression of eNOS and alpha(v)beta(3) integrin and between glandular expression of eNOS and luminal expression of alpha(v)beta(3) integrin (PMID:12372469)
• activity decreases during type I collagen-induced platelet aggregation and may be controlled with differential phosphorylation of specific amino acid residues located in the enzyme (PMID:12379480)
• While eNOS seems to be involved in the physiological regulation of hepatic perfusion in fulminant hepatic failure (PMID:12399227)
• Polymorphism of this gene is not related to the development of diabetic nephropathy. (PMID:12413777)
• Association analysis of five polymorphic microsatellite markers in cluster headache (CH) patients reveals that genetic variations in NOS3 do not contribute greatly to CH susceptibility. (PMID:12421162)
• regression analyses indicate no substantial effect of G894T polymorphism on the variance of plasma NO(x) levels in healthy Korean population (PMID:12431478)
• The (-)786T-C mutation of the eNOS gene is associated with insulin resistance in both Japanese non-diabetic subjects and Type II diabetic patients (PMID:12436344)
• nitric oxide synthases(inducible, brain and endothelial) were expressed in human pregnant term uterus and did not change in the myometrium during labor (PMID:12445599)
• we have identified a nitric oxide related genetic factor associated with micrometastization of prostate cancer (PMID:12445681)
• data demonstrate that Rho/ROCK pathway negatively regulates eNOS phosphorylation through inhibition of protein kinase B (PKB), whereas it downregulates eNOS expression independent of PKB. (PMID:12446767)
• The G894T mutation of the endothelial nitric oxide synthase gene may be a marker for genetical predisposition of CHD in Chinese Han population. (PMID:12476417)
• Tandem repeat polymorphism in intron 4 of Nos3 was analyzed by polymerase chain reaction. The common B allele was identified on 143 of 182 chromosomes (frequency 0.79). The polymorphic A allele was present on 39 chromosomes (frequency 0.21). (PMID:12477509)

Cross-species orthologs

2 orthologs

Organism	Symbol	Gene ID
mus_musculus	Nos3	ENSMUSG00000028978
rattus_norvegicus	Nos3	ENSRNOG00000009348

Paralogs (5): NOS2 (ENSG00000007171), NOS1 (ENSG00000089250), MTRR (ENSG00000124275), POR (ENSG00000127948), NDOR1 (ENSG00000188566)

Mouse-model phenotypes

Observed phenotypes of the mouse ortholog (MGI:97362) from MGI (Alliance of Genome Resources) — knockout/mutant phenotypes, ranked by annotation count. 225 distinct phenotype terms.

Mouse phenotype (MP)	Records
hypertension	7
decreased heart rate	6
increased systemic arterial systolic blood pressure	5
abnormal systemic arterial blood pressure	4
decreased body weight	4
increased mean systemic arterial blood pressure	4
increased circulating triglyceride level	3
increased heart rate	3
increased systemic arterial blood pressure	3
increased vasoconstriction	3
decreased cardiac muscle contractility	3
abnormal renal glomerulus morphology	3
insulin resistance	3
decreased vasodilation	3
abnormal heart right ventricle pressure	3
decreased superovulation rate	3
decreased bone mineral density	2
abnormal blood vessel physiology	2
abnormal heart morphology	2
small ovary	2
increased circulating free fatty acids level	2
abnormal vasodilation	2
cardiac hypertrophy	2
decreased litter size	2
increased circulating insulin level	2

+200 more (showing top 25).

Additional cross-species homologs

Sequence homologs in species beyond Ensembl Compara’s model organisms, by Diamond local protein alignment (% sequence identity; best hit per species). Lower-identity rows are more distant homologs. The matched accessions’ gene symbols aren’t in biobtree yet, so each row shows the UniProt accession.

Organism	UniProt	Identity	Method
Staphylococcus aureus	P0A004	100.0%	Diamond
Staphylococcus aureus subsp. aureus Mu50	P0A092	100.0%	Diamond
Staphylococcus aureus subsp. aureus N315	P0A093	100.0%	Diamond
Staphylococcus aureus subsp. aureus MW2	P0A094	100.0%	Diamond
Staphylococcus aureus subsp. aureus COL	Q5HEK7	100.0%	Diamond
Staphylococcus aureus subsp. aureus MSSA476	Q6G818	100.0%	Diamond
Staphylococcus aureus subsp. aureus MRSA252	Q6GFE2	98.6%	Diamond
Bos taurus	Q27995	97.0%	Diamond
Capra hircus	Q28314	97.0%	Diamond
Vicia faba	P41346	96.7%	Diamond
Sus scrofa	Q28969	95.6%	Diamond
Oryctolagus cuniculus	O19114	95.4%	Diamond
Cavia porcellus	P97270	94.6%	Diamond
Canis lupus familiaris	Q9TUX8	93.5%	Diamond
Sorghum bicolor	C5YJG8	92.8%	Diamond
Ovis aries	Q29498	91.9%	Diamond
Squalus acanthias	Q9I9M2	88.2%	Diamond
Spinacia oleracea	P00455	86.7%	Diamond
Arabidopsis thaliana	Q9FKW6	85.0%	Diamond
Oryza sativa Japonica Group	Q653S9	80.2%	Diamond
Artemisia annua	A0A2U1LIM9	79.7%	Diamond
Spirulina sp.	P00454	74.9%	Diamond
Synechocystis sp. PCC 6803 substr. Kazusa	Q55318	74.9%	Diamond
Thermosynechococcus vestitus BP-1	Q93RE3	72.8%	Diamond
Anopheles stephensi	O61608	71.2%	Diamond
Drosophila melanogaster	Q27571	71.2%	Diamond
Carassius auratus	Q92037	70.4%	Diamond
Oncorhynchus mykiss	Q92091	70.4%	Diamond
Gallus gallus	Q90703	69.4%	Diamond
Bombyx mori	B1B557	65.5%	Diamond
Rhodnius prolixus	Q26240	62.6%	Diamond
Bacillus subtilis subsp. subtilis str. 168	O08336	60.7%	Diamond
Lymnaea stagnalis	O61309	47.9%	Diamond
Schizosaccharomyces pombe 972h-	Q09744	47.2%	Diamond
Deinococcus radiodurans R1 = ATCC 13939 = DSM 20539	Q9RR97	42.5%	Diamond

Genomic neighbors (locus context)

Genes flanking this one on the chromosome (NCBI/Entrez gene neighborhood). Positional context only — some neighbors share regulatory elements (bidirectional promoters, antisense/overlapping transcripts), but chromosomal adjacency is not itself a functional relationship (see Interactions for that).

Symbol	Type	Entrez
AOC1	protein-coding	26
ATG9B	protein-coding	285973
TMEM176B	protein-coding	28959
KCNH2	protein-coding	3757
TMEM176A	protein-coding	55365
CDK5	protein-coding	1020
ASIC3	protein-coding	9311
ABCB8	protein-coding	11194

Protein

Protein identifiers

Nitric oxide synthase 3 — P29474 (reviewed: P29474)

Alternative names: Constitutive NOS, EC-NOS, NOS type III, Nitric oxide synthase, endothelial

All UniProt accessions (3): P29474, E7ESA7, H7C4V4

UniProt curated annotations — full annotation on UniProt →

Function. Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets. Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.

Subunit / interactions. Homodimer. Interacts with NOSIP and NOSTRIN. Interacts with HSP90AB1. Forms a complex with ASL, ASS1 and SLC7A1; the complex regulates cell-autonomous L-arginine synthesis and citrulline recycling while channeling extracellular L-arginine to nitric oxide synthesis pathway.

Subcellular location. Cell membrane. Membrane. Caveola. Cytoplasm. Cytoskeleton. Golgi apparatus.

Tissue specificity. Platelets, placenta, liver and kidney.

Post-translational modifications. Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity. Phosphorylation of Ser-114 by CDK5 reduces activity.

Disease relevance. Moyamoya disease 8 (MYMY8) [MIM:621469] A form of Moyamoya disease, a progressive cerebral angiopathy characterized by bilateral intracranial carotid artery stenosis and telangiectatic vessels in the region of the basal ganglia. The abnormal vessels resemble a ‘puff of smoke’ (moyamoya) on cerebral angiogram. Affected individuals can develop transient ischemic attacks and/or cerebral infarction, and rupture of the collateral vessels can cause intracranial hemorrhage. Hemiplegia of sudden onset and epileptic seizures constitute the prevailing presentation in childhood, while subarachnoid bleeding occurs more frequently in adults. MYMY8 inheritance is autosomal recessive. The disease may be caused by variants affecting the gene represented in this entry.

Activity regulation. Stimulated by calcium/calmodulin. Inhibited by NOSIP and NOSTRIN.

Cofactor. Binds 1 FAD. Binds 1 FMN. Tetrahydrobiopterin (BH4). May stabilize the dimeric form of the enzyme.

Miscellaneous. Lacks eNOS activity.

Similarity. Belongs to the NOS family.

Isoforms (3)

UniProt ID	Names	Canonical?
P29474-1	1	yes
P29474-2	eNOS13C
P29474-3	eNOS13B

RefSeq proteins (4): NP_000594, NP_001153581, NP_001153582, NP_001153583 (=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR001094	Flavdoxin-like	Domain
IPR001433	OxRdtase_FAD/NAD-bd	Domain
IPR001709	Flavoprot_Pyr_Nucl_cyt_Rdtase	Domain
IPR003097	CysJ-like_FAD-binding	Domain
IPR004030	NOS_N	Domain
IPR008254	Flavodoxin/NO_synth	Domain
IPR012144	NOS_euk	Family
IPR017927	FAD-bd_FR_type	Domain
IPR017938	Riboflavin_synthase-like_b-brl	Homologous_superfamily
IPR023173	NADPH_Cyt_P450_Rdtase_alpha	Homologous_superfamily
IPR029039	Flavoprotein-like_sf	Homologous_superfamily
IPR036119	NOS_N_sf	Homologous_superfamily
IPR039261	FNR_nucleotide-bd	Homologous_superfamily
IPR044940	NOS_dom_2	Homologous_superfamily
IPR044943	NOS_dom_1	Homologous_superfamily
IPR044944	NOS_dom_3	Homologous_superfamily
IPR050607	NOS	Family

Pfam: PF00175, PF00258, PF00667, PF02898

Enzyme classification (BRENDA)

• EC 1.14.13.39 — nitric-oxide synthase (NADPH) (BRENDA: 31 organisms, 140 substrates, 179 inhibitors, 88 Km, 25 kcat entries)

Substrate kinetics (BRENDA)

11 substrates with measured Km, best-characterized 11. Km ranges are aggregated across organisms/conditions.

Substrate	Km (mM)	Measurements
L-ARGININE	0.0019–68.5	28
NADPH	0.0003–0.0041	9
TETRAHYDROBIOPTERIN	—	6
ADRIAMYCIN	0.012–0.078	5
MENADIONE	0.01–0.041	5
MITOMYCIN C	0.0073–0.055	5
NGAMMA-HYDROXY-L-ARGININE	0.019–0.129	5
2’,3’-DIALDEHYDE-NADPH	0.0008	1
CALMODULIN	—	1
NITROBLUE TETRAZOLIUM	0.016	1
NITRIC OXIDE	—	0

Catalyzed reactions (Rhea)

• 2 L-arginine + 3 NADPH + 4 O2 + H(+) = 2 L-citrulline + 2 nitric oxide + 3 NADP(+) + 4 H2O (RHEA:19897)

UniProt features (137 total): binding site 42, helix 27, strand 23, modified residue 10, sequence variant 9, sequence conflict 7, splice variant 4, region of interest 3, lipid moiety-binding region 3, turn 2, domain 2, compositionally biased region 2, initiator methionine 1, chain 1, mutagenesis site 1

Structure

Experimental structures (PDB)

showing 30 of 105 structures by resolution:

PDB	Title	Method	Resolution (Å)
9CWI	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.73
4D1P	Structure of human endothelial nitric oxide synthase heme domain IN COMPLEX WITH …	X-RAY DIFFRACTION	1.73
9MWU	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.74
6PP1	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.76
9MWO	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.76
8UFT	Structure of human endothelial nitric oxide synthase P370N mutant heme domain in complex …	X-RAY DIFFRACTION	1.78
8FGO	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.8
6NH8	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.8
9Q51	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.8
9MWW	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.8
4D1O	Structure of human endothelial nitric oxide synthase heme domain with L-Arg bound	X-RAY DIFFRACTION	1.82
6NHF	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.83
9CW7	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.83
9MWX	Structure of human endothelial nitric oxide synthase heme domain bound …	X-RAY DIFFRACTION	1.84
8FGS	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.84
7TSG	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.85
7TSM	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.85
9MWP	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.86
8UFR	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.87
8FGP	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.88
9CW5	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.88
9Q53	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.88
9MWV	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.9
9MWS	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.9
6NH7	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.9
7TSL	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.9
7UAO	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.9
8FGT	Structure of human endothelial nitric oxide synthase heme domain in complex with …	X-RAY DIFFRACTION	1.9
9CWJ	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.9
9CWK	Structure of human endothelial nitric oxide synthase heme domain bound with …	X-RAY DIFFRACTION	1.9

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-P29474-F1	83.20	0.56

Human Protein Atlas

Protein class: Enzymes, ENZYME proteins, Oxidoreductases, Metabolic proteins, Predicted intracellular proteins, Plasma proteins, Cancer-related genes, Candidate cancer biomarkers, Disease related genes, FDA approved drug targets, Small molecule drugs, Human disease related genes

Protein evidence: Evidence at protein level

Antibody reliability (HPA): IH enhanced

Functional residue map

Curated UniProt residues grouped by drug-discovery relevance — catalytic, ligand-binding, modification, and mutation-validated positions. Source: UniProtKB sequence features.

Ligand- & substrate-binding residues (42)

Position	Role
99	Zn(2+)
102	(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
184	heme b
247	L-arginine
356	L-arginine
357	L-arginine
361	L-arginine
365	(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
366	L-arginine
446	(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
447	(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
460	(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
475	heme b
526	FMN
527	FMN
528	FMN
530	FMN
572	FMN
573	FMN
654	FMN
661	FMN
687	FMN
691	FMN
776	NADP(+)
798	FAD
938	FAD
940	FAD
941	FAD
956	FAD
958	FAD
962	FAD
975	FAD
976	FAD
977	FAD
1016	NADP(+)
1049	NADP(+)
1078	NADP(+)
1079	NADP(+)
1085	NADP(+)
1087	NADP(+)
1089	NADP(+)
94	Zn(2+)

Post-translational modifications (13)

Type	Positions
Phosphothreonine	33, 495, 1175
Phosphoserine	114, 615, 633, 638, 836, 1177, 1179
N-myristoyl glycine	2
S-palmitoyl cysteine	15, 26

Mutagenesis-validated functional residues (1)

Position	Phenotype
114	reduced nitrite (no) production.

Function

Pathways and Gene Ontology

Reactome pathways

8 curated (TAS) of 26; 0 disease pathways. Curated-first; evidence = TAS (manually curated) vs IEA (electronic inference).

26 pathways:

ID	Pathway	Evidence
R-HSA-9009391	Extra-nuclear estrogen signaling	curated
R-HSA-9856530	High laminar flow shear stress activates signaling by PIEZO1 and PECAM1:CDH5:KDR in endothelial cells	curated
R-HSA-203754	NOSIP mediated eNOS trafficking	curated
R-HSA-203641	NOSTRIN mediated eNOS trafficking	curated
R-HSA-392154	Nitric oxide stimulates guanylate cyclase	curated
R-HSA-1222556	ROS and RNS production in phagocytes	curated
R-HSA-1474151	Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation	curated
R-HSA-5218920	VEGFR2 mediated vascular permeability	curated
R-HSA-9855142	Cellular responses to mechanical stimuli	—
R-HSA-8953897	Cellular responses to stimuli	—
R-HSA-8939211	ESR-mediated signaling	—
R-HSA-109582	Hemostasis	—
R-HSA-168256	Immune System	—
R-HSA-168249	Innate Immune System	—
R-HSA-1430728	Metabolism	—
R-HSA-8978934	Metabolism of cofactors	—
R-HSA-202131	Metabolism of nitric oxide: NOS3 activation and regulation	—
R-HSA-196854	Metabolism of vitamins and cofactors	—
R-HSA-418346	Platelet homeostasis	—
R-HSA-9860931	Response of endothelial cells to shear stress	—
R-HSA-162582	Signal Transduction	—
R-HSA-9006931	Signaling by Nuclear Receptors	—
R-HSA-9006934	Signaling by Receptor Tyrosine Kinases	—
R-HSA-194138	Signaling by VEGF	—
R-HSA-4420097	VEGFA-VEGFR2 Pathway	—
R-HSA-203615	eNOS activation	electronic

MSigDB gene sets: 446 (showing top): PID_SHP2_PATHWAY, GOBP_CARDIAC_CHAMBER_DEVELOPMENT, GOBP_POTASSIUM_ION_TRANSPORT, BIOCARTA_GCR_PATHWAY, GOBP_REGULATION_OF_CELL_ACTIVATION, GOBP_EMBRYO_DEVELOPMENT_ENDING_IN_BIRTH_OR_EGG_HATCHING, REACTOME_INNATE_IMMUNE_SYSTEM, GOBP_CARDIAC_SEPTUM_DEVELOPMENT, GOBP_REGULATION_OF_BLOOD_PRESSURE, GOBP_VENTRICULAR_SEPTUM_MORPHOGENESIS, GOBP_NEGATIVE_REGULATION_OF_SMOOTH_MUSCLE_CELL_PROLIFERATION, GOBP_REGULATION_OF_WOUND_HEALING, GOBP_CIRCULATORY_SYSTEM_PROCESS, GOBP_INTRACELLULAR_PROTEIN_TRANSPORT, GOMF_OXIDOREDUCTASE_ACTIVITY_ACTING_ON_PAIRED_DONORS_WITH_INCORPORATION_OR_REDUCTION_OF_MOLECULAR_OXYGEN

GO Biological Process (35): blood vessel remodeling (GO:0001974), regulation of systemic arterial blood pressure by endothelin (GO:0003100), aortic valve morphogenesis (GO:0003180), pulmonary valve morphogenesis (GO:0003184), endocardial cushion morphogenesis (GO:0003203), L-arginine catabolic process (GO:0006527), nitric oxide biosynthetic process (GO:0006809), mitochondrion organization (GO:0007005), regulation of blood pressure (GO:0008217), negative regulation of cell population proliferation (GO:0008285), response to heat (GO:0009408), response to hormone (GO:0009725), negative regulation of platelet activation (GO:0010544), positive regulation of gene expression (GO:0010628), negative regulation of muscle hyperplasia (GO:0014740), smooth muscle hyperplasia (GO:0014806), removal of superoxide radicals (GO:0019430), regulation of nervous system process (GO:0031644), response to lipopolysaccharide (GO:0032496), response to fluid shear stress (GO:0034405), vasodilation (GO:0042311), positive regulation of blood vessel endothelial cell migration (GO:0043536), endothelial cell migration (GO:0043542), cell redox homeostasis (GO:0045454), positive regulation of Notch signaling pathway (GO:0045747), positive regulation of angiogenesis (GO:0045766), negative regulation of blood pressure (GO:0045776), tetrahydrobiopterin metabolic process (GO:0046146), nitric oxide metabolic process (GO:0046209), homeostasis of number of cells within a tissue (GO:0048873), ventricular septum morphogenesis (GO:0060412), negative regulation of biomineral tissue development (GO:0070168), blood vessel diameter maintenance (GO:0097746), negative regulation of extrinsic apoptotic signaling pathway via death domain receptors (GO:1902042), blood circulation (GO:0008015)

GO Molecular Function (15): actin monomer binding (GO:0003785), nitric-oxide synthase activity (GO:0004517), calmodulin binding (GO:0005516), FMN binding (GO:0010181), superoxide-generating NAD(P)H oxidase activity (GO:0016175), heme binding (GO:0020037), tetrahydrobiopterin binding (GO:0034617), arginine binding (GO:0034618), cadmium ion binding (GO:0046870), flavin adenine dinucleotide binding (GO:0050660), NADP binding (GO:0050661), scaffold protein binding (GO:0097110), protein binding (GO:0005515), oxidoreductase activity (GO:0016491), metal ion binding (GO:0046872)

GO Cellular Component (9): Golgi membrane (GO:0000139), nucleus (GO:0005634), cytoplasm (GO:0005737), Golgi apparatus (GO:0005794), cytosol (GO:0005829), cytoskeleton (GO:0005856), plasma membrane (GO:0005886), caveola (GO:0005901), endocytic vesicle membrane (GO:0030666)

Reactome top-level categories

13 top-level categories:

Category	Pathways
Metabolism of nitric oxide: NOS3 activation and regulation	3
Metabolism	2
Innate Immune System	1
Metabolism of cofactors	1
Platelet homeostasis	1
VEGFA-VEGFR2 Pathway	1
ESR-mediated signaling	1
Response of endothelial cells to shear stress	1
Immune System	1
Signaling by Receptor Tyrosine Kinases	1
Hemostasis	1
Signaling by VEGF	1
Signaling by Nuclear Receptors	1

GO top-level categories

showing 60 of 70 GO namespaces:

Category	Terms
heart valve morphogenesis	2
response to stress	2
muscle hyperplasia	2
protein binding	2
anion binding	2
binding	2
cation binding	2
bounding membrane of organelle	2
intracellular membrane-bounded organelle	2
cellular anatomical structure	2
cytoplasm	2
tissue remodeling	1
regulation of systemic arterial blood pressure by hormone	1
aortic valve development	1
pulmonary valve development	1
heart morphogenesis	1
endocardial cushion development	1
mesenchyme morphogenesis	1
arginine metabolic process	1
amino acid catabolic process	1
proteinogenic amino acid catabolic process	1
biosynthetic process	1
nitric oxide metabolic process	1
organelle organization	1
blood circulation	1
regulation of biological quality	1
cell population proliferation	1
regulation of cell population proliferation	1
negative regulation of cellular process	1
response to temperature stimulus	1
response to endogenous stimulus	1
response to chemical	1
regulation of platelet activation	1
platelet activation	1
negative regulation of blood coagulation	1
negative regulation of cell activation	1
gene expression	1
regulation of gene expression	1
positive regulation of macromolecule biosynthetic process	1
regulation of muscle hyperplasia	1
negative regulation of muscle adaptation	1
smooth muscle adaptation	1
superoxide metabolic process	1
cellular response to superoxide	1
cellular oxidant detoxification	1
regulation of system process	1
nervous system process	1
response to molecule of bacterial origin	1
response to lipid	1
response to oxygen-containing compound	1
actin binding	1
oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen	1
ribonucleotide binding	1
oxidoreductase activity, acting on NAD(P)H, oxygen as acceptor	1
tetrapyrrole binding	1
amino acid binding	1
carboxylic acid binding	1
transition metal ion binding	1
nucleotide binding	1
adenyl nucleotide binding	1

Experimentally-supported GO annotations

GO terms backed by direct experimental evidence (ECO experimental codes) — 18 of 60 annotations. The rest are phylogenetic / computational / electronic (IEA).

18 experimentally-supported GO terms:

GO term	NS	Evidence
L-arginine catabolic process	BP	direct assay
endothelial cell migration	BP	mutant phenotype
nitric oxide biosynthetic process	BP	direct assay
positive regulation of angiogenesis	BP	direct assay
positive regulation of blood vessel endothelial cell migration	BP	direct assay
regulation of systemic arterial blood pressure by endothelin	BP	mutant phenotype
removal of superoxide radicals	BP	direct assay
response to fluid shear stress	BP	expression pattern
Golgi apparatus	CC	direct assay
caveola	CC	direct assay
cytoplasm	CC	direct assay
plasma membrane	CC	direct assay
actin monomer binding	MF	physical interaction
arginine binding	MF	direct assay
calmodulin binding	MF	experiment
heme binding	MF	direct assay
nitric-oxide synthase activity	MF	direct assay
tetrahydrobiopterin binding	MF	direct assay

Protein interactions and networks

STRING

showing 15 of 2,920 interactions by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
NOS3	EDN1	P05305	867
NOS3	AKT1	P31749	848
NOS3	DDAH1	O94760	840
NOS3	XDH	P47989	821
NOS3	IL1B	P01584	783
NOS3	IL6	P05231	780
NOS3	AGT	P01019	774
NOS3	CALM1	P02593	767
NOS3	CALML3	P27482	766
NOS3	CALML6	Q8TD86	766
NOS3	CALML4	Q96GE6	766
NOS3	CALML5	Q9NZT1	766
NOS3	ACTB	P02570	762
NOS3	INS	P01308	741
NOS3	PTGS2	P35354	740

IntAct

Human interaction partners of NOS3 in IntAct, one row per partner (best MIscore; records = supporting IntAct interaction records).

showing 30 of 84 partners by confidence:

Partner	Interaction type	MIscore	Records
NOSTRIN	physical association	0.730	9
GOLGA2	physical association	0.720	6
CDC37	direct interaction	0.580	4
AP2B1	physical association	0.560	3
ARFGAP1	physical association	0.560	3
ASB3	physical association	0.560	3
ASIC4	physical association	0.560	3
C6orf141	physical association	0.560	3
COPS3	physical association	0.560	3
CRLF3	physical association	0.560	3
FANCG	physical association	0.560	3
H3C13	physical association	0.560	3
HNRNPK	physical association	0.560	3
HYAL2	physical association	0.560	3
IKZF3	physical association	0.560	3
KLHL20	physical association	0.560	3
KRT33B	physical association	0.560	3
LGALS9C	physical association	0.560	3
MAGEA11	physical association	0.560	3
METTL27	physical association	0.560	3
MICAL2	physical association	0.560	3
MKL1	physical association	0.560	3
MPND	physical association	0.560	3
NSMF	physical association	0.560	3
NVL	physical association	0.560	3
PCMTD2	physical association	0.560	3
PCSK7	physical association	0.560	3
PDZK1	physical association	0.560	3
PIAS1	physical association	0.560	3
PPP1R16A	physical association	0.560	3

BioGRID (108 interactions; top human partners by supporting records): HSP90AA1 (Affinity Capture-Western; 10 records), CALM1 (Reconstituted Complex; 7 records), CAV1 (Affinity Capture-Western; 6 records), NOSIP (Affinity Capture-Western; 6 records), NOSTRIN (Two-hybrid; 6 records), AKT1 (Biochemical Activity; 5 records), ACTB (Affinity Capture-Western; 4 records), CDC37 (Affinity Capture-Western; 3 records), GUCY1B3 (Reconstituted Complex; 3 records), APOE (Two-hybrid; 2 records), APP (Affinity Capture-Western; 2 records), GCDH (Affinity Capture-Western; 2 records), GOLGA2 (Two-hybrid; 2 records), PPP2R4 (Affinity Capture-Western; 2 records), ST13 (Affinity Capture-Western; 2 records)

SIGNOR signaling

showing 41 of 45 signaling interactions:

A	Effect	B	Mechanism
AKT1	up-regulates	NOS3	phosphorylation
PRKAB1	up-regulates	NOS3	phosphorylation
PRKACA	up-regulates	NOS3	phosphorylation
PRKACG	up-regulates	NOS3	phosphorylation
PRKAA1	up-regulates	NOS3	phosphorylation
PTK2B	down-regulates	NOS3	phosphorylation
CDK5	down-regulates	NOS3	phosphorylation
AMPK	up-regulates	NOS3	phosphorylation
AKT	up-regulates	NOS3	phosphorylation
PPP1CC	“up-regulates activity”	NOS3	dephosphorylation
PPP1CA	“up-regulates activity”	NOS3	dephosphorylation
PPP1CB	“up-regulates activity”	NOS3	dephosphorylation
CALM1	“up-regulates activity”	NOS3	binding
PRKACA	“up-regulates activity”	NOS3	phosphorylation
AMPK	“down-regulates activity”	NOS3	phosphorylation
PRKAA1	“up-regulates activity”	NOS3	phosphorylation
PRKCA	“down-regulates activity”	NOS3	phosphorylation
AKT1	“up-regulates activity”	NOS3	phosphorylation
AKT2	“up-regulates activity”	NOS3	phosphorylation
AKT3	“up-regulates activity”	NOS3	phosphorylation
AKT	“up-regulates activity”	NOS3	phosphorylation
NOS3	“up-regulates quantity”	“nitric oxide”	“chemical modification”
PRKCB	“down-regulates activity”	NOS3	phosphorylation
PRKCD	“down-regulates activity”	NOS3	phosphorylation
PRKCE	“down-regulates activity”	NOS3	phosphorylation
PRKCG	“down-regulates activity”	NOS3	phosphorylation
PRKCH	“down-regulates activity”	NOS3	phosphorylation
PRKCI	“down-regulates activity”	NOS3	phosphorylation
PRKCQ	“down-regulates activity”	NOS3	phosphorylation
PRKCZ	“down-regulates activity”	NOS3	phosphorylation
HSP90AB1	“up-regulates activity”	NOS3	binding
PP1	“up-regulates activity”	NOS3	dephosphorylation
CALM2	“up-regulates activity”	NOS3	binding
CALM3	“up-regulates activity”	NOS3	binding
PKC	“down-regulates activity”	NOS3	phosphorylation
PTPRB	“down-regulates activity”	NOS3	dephosphorylation
ABL1	“up-regulates activity”	NOS3	phosphorylation
SRC	“up-regulates activity”	NOS3	phosphorylation
EPHB4	“up-regulates activity”	NOS3	phosphorylation
HSP90AA1	up-regulates	NOS3	binding
CRP	“down-regulates quantity by destabilization”	NOS3

Protein complexes (CORUM) (5)

Experimentally-characterized complexes this protein is a subunit of (★ = the complex contains a known drug target).

5 complexes by size:

Complex	Subunits	Members
UTM-SGCE-DAG1-CAV1-NOS3 complex	5	CAV1, DAG1, NOS3, SGCE, UTRN
NOS3-CAV1-NOSTRIN complex	3	CAV1, NOS3, NOSTRIN
★ AKT1-HSP90AB1-NOS3 complex	3	HSP90AB1, NOS3, AKT1
NOS3-CAV1 complex	2	CAV1, NOS3
★ HSP90AB1-NOS3 complex	2	HSP90AB1, NOS3

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 84 IntAct physical interaction partners (hypergeometric vs the genome-wide background, BH-FDR, gene-set size 15–500, ranked by fold). A functional readout of the neighbourhood — distinct from this gene’s own memberships above, and biased toward well-studied / hub proteins, so read it as themes rather than proof.

Reactome pathways:

Pathway	Partners	Fold	FDR
Oncogenic MAPK signaling	6	25.7×	7e-05
MAPK1/MAPK3 signaling	6	13.6×	7e-04
MAPK family signaling cascades	6	10.6×	2e-03
Diseases of signal transduction by growth factor receptors and second messengers	9	8.8×	2e-04
Signaling by Receptor Tyrosine Kinases	9	8.0×	3e-04
RAF/MAP kinase cascade	7	7.4×	3e-03

GO biological processes:

GO term	Partners	Fold	FDR
epidermal growth factor receptor signaling pathway	6	20.1×	6e-04

Disease & clinical

Genetic variants

Variant evidence for NOS3: 191 ClinVar records (4 pathogenic / likely pathogenic). Per-variant detail is on Sugi Variant.

ClinVar

191 variants total. Per-class counts are floors (≥ shown; pagination cap):

Classification	Count (floor)
Pathogenic	2
Likely pathogenic	2
Uncertain significance	109
Likely benign	29
Benign	33

Top pathogenic / likely-pathogenic (4)

Showing all 4 pathogenic / likely-pathogenic ClinVar records, ranked by ClinVar review status (expert panel → multiple submitters → single submitter), then by number of submitting labs.

Variant ID	HGVS	Classification	Review	Submitters
929757	NM_000603.5(NOS3):c.172C>T (p.Pro58Ser)	Likely pathogenic	criteria provided, single submitter	1
929758	NM_000603.5(NOS3):c.505G>A (p.Glu169Lys)	Likely pathogenic	criteria provided, single submitter	1
4686577	NOS3, IVS12, G-C, +1	Pathogenic	no assertion criteria provided	1
4686578	C648R	Pathogenic	no assertion criteria provided	1

Per-variant reference — Sugi Variant

Explore NOS3’s pathogenic and likely-pathogenic variants individually — each with its ClinVar classification and conditions, applied ACMG criteria (ClinGen expert panel, where curated), the calibrated in-silico predictors (AlphaMissense, REVEL, SaProt, conservation), gnomAD frequency, and the predictor-disagreement QC signal — on Sugi Variant.

AlphaMissense

3,281 missense substitutions predicted likely pathogenic (of 7,770 scored), across 776 residues. Top 20 hotspot residues, ranked by the number of likely-pathogenic substitutions (of those reachable by a single-nucleotide change), then by mean score:

Residue	Likely-pathogenic substitutions	Mean am_pathogenicity	Max am_pathogenicity
M1119	9	0.956	0.996
M207	9	0.855	0.993
D1158	8	0.997	0.999
H798	8	0.994	0.997
D264	8	0.990	0.997
F1160	8	0.990	1.000
D82	8	0.989	0.997
D396	8	0.989	0.997
H214	8	0.988	0.997
D1054	8	0.987	0.995
F231	8	0.986	0.998
F546	8	0.986	0.997
D686	8	0.986	0.997
F999	8	0.985	0.998
D1116	8	0.985	0.997
D419	8	0.981	0.997
H461	8	0.980	0.997
F650	8	0.980	0.998
F353	8	0.979	0.998
D864	8	0.979	0.993

HPA cancer prognostics

RNA cancer specificity: Low cancer specificity

Human Protein Atlas survival analysis — favorable = higher expression predicts better survival.

3 prognostic cancer types:

Cancer	Prognostic	p-value
Kidney Renal Clear Cell Carcinoma (TCGA)	favorable	3.84e-06
Kidney Renal Papillary Cell Carcinoma (TCGA)	unfavorable	2.68e-05
Liver Hepatocellular Carcinoma (validation)	favorable	0.000324

Disease associations

OMIM: gene MIM:163729 | disease phenotypes: MIM:621469

Genomics England panels (PanelApp)

Clinical diagnostic gene panels including this gene (1 panels, 0 green/diagnostic-grade). Confidence: green = diagnostic, amber = moderate.

1 panels:

Panel	Rating
Cerebral vascular malformations	amber

Mondo (2): Moyamoya disease 8 (MONDO:0980949), primary ovarian failure (MONDO:0005387)

Orphanet (1): NON RARE IN EUROPE: Primary ovarian failure (Orphanet:619)

MeSH disease descriptors

1 disease descriptors:

Descriptor	Name	Tree numbers
D016649	Primary Ovarian Insufficiency	C12.050.351.500.056.630.750; C12.100.250.056.630.750; C19.391.630.750

Drugs & pharmacology

Drug and pharmacology data

Is drug target: yes

ChEMBL targets (4): CHEMBL2111350 (SELECTIVITY GROUP), CHEMBL2111405 (SELECTIVITY GROUP), CHEMBL4630725 (PROTEIN FAMILY), CHEMBL4803 (SINGLE PROTEIN)

Drugs acting on a complex or family that includes this protein

Curated ChEMBL mechanism-of-action drugs whose target is a protein complex or family of ≤10 members that includes NOS3 — the drug acts on the assembly or class, not necessarily on this subunit alone.

1 drugs via a complex or family:

Drug	Target (ChEMBL)	Members	Max phase
TILARGININE	Nitric oxide synthase	3	3

Molecules with ChEMBL bioactivity

Phase ≥1, by development phase (incl. off-target/promiscuous compounds). Patent mentions across the top 20 by phase: 48,254 (via chembl_molecule»patent_compound — counts attach to the compound, not the gene–compound relationship, so off-target/promiscuous molecules can dominate).

6 molecules by phase:

Molecule	Name	Phase	Patents
CHEMBL1371	CHLORZOXAZONE	4	16,875
CHEMBL256147	TILARGININE	3	2,046
CHEMBL114551	GW-274150	2	1,071
CHEMBL7890	L-NAME	2	1,196
CHEMBL225304	PIMAGEDINE	2	24,594
CHEMBL227744	NITROARGININE	1	2,472

PharmGKB: 1 entry (CPIC guideline: no)

GtoPdb / IUPHAR curated pharmacology

(IUPHAR/BPS Guide to Pharmacology — expert-curated)

Target class: enzyme — Nitric oxide synthases

Most potent curated ligand interactions (3 total), top 3:

Ligand	Action	Affinity	Parameter
L-NNA	Inhibition	7.52	pKi
tilarginine	Inhibition	6.4	pKi
GW274150	Inhibition	4.47	pIC50

Binding affinities (BindingDB)

774 measured of 913 human assays (1005 total across all organisms); most potent 50 below. Values come from heterogeneous assays and are not directly comparable.

Ligand	Measure	Value	Patent
CHEMBL2046871	KI	0.43 nM
CHEMBL106871	IC50	1 nM
6-{4-[2-(4-Quinolin-8-yl-piperazin-1-yl)-ethyl]-phenyl}-pyridin-2-ylamine	KI	3 nM
N-(3-Aminomethyl-phenyl)-furan-2-carboxamidine	KI	6 nM
N-(3-Aminomethyl-phenyl)-furan-2-carboxamidine	KI	6.3 nM
N-(3-Aminomethyl-phenyl)-thiophene-2-carboxamidine	KI	8.7 nM
4-Methyl-6-propylpyridin-2-amine, 7	IC50	10 nM
6-[[(2R,4S)-4-[(6-amino-4-methyl-2-pyridinyl)methoxy]pyrrolidin-2-yl]methoxymethyl]-4-methylpyridin-2-amine	KI	10 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
N-(3-Aminomethyl-phenyl)-2-fluoro-acetamidine	KI	11 nM
N-(3-Aminomethyl-phenyl)-2-methylsulfanyl-acetamidine	KI	11 nM
CHEMBL1944717	IC50	12 nM
CHEMBL2046877	KI	12 nM
N’-[4-[3-(dimethylamino)propyl]-2,3-dihydro-1,4-benzothiazin-7-yl]thiophene-2-carboximidamide	IC50	14 nM	US-8618286: Benzoxazines, benzothiazines, and related compounds having NOS inhibitory activity
4-Methyl-piperidin-(2Z)-ylideneamine	IC50	16 nM
N1-((3R,4S)-4-((6-aminopyridin-2-yl)methyl)pyrrolidin-3-yl)-N2-(4-fluorobenzyl)ethane-1,2-diamine tetrahydrochloride	KI	17.7 nM	US-9090589: Specific nNOS inhibitors for the therapy and prevention of human melanoma
ONO-1714	KI	18.6 nM
CHEMBL5420690	KI	19 nM
CHEMBL4068062	KI	20 nM
CHEMBL1825174	IC50	20 nM
CHEMBL4089246	KI	21 nM
CHEMBL4532683	KI	21 nM
CHEMBL5191295	KI	21 nM
CHEMBL4590333	KI	23 nM
6-[2-[5-[(2R)-1-amino-3-(6-amino-4-methyl-2-pyridinyl)propan-2-yl]-3-pyridinyl]ethyl]-4-methylpyridin-2-amine	KI	24 nM	US-9951014: Mammalian and bacterial nitric oxide synthase inhibitors
N1-{(+/-)-4’-[(6’’-amino-4’’-methylpyridin-2’’-yl)methyl]pyrrolidin-3’-yl}-N2-phenethylethane-1,2-diamine tetrahydrochloride	KI	24 nM	US-9090589: Specific nNOS inhibitors for the therapy and prevention of human melanoma
6-[[(2S,4R)-4-[(6-amino-4-methyl-2-pyridinyl)methoxy]pyrrolidin-2-yl]methoxymethyl]-4-methylpyridin-2-amine	KI	26 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
N’-[4-[3-(methylamino)propyl]-2,3-dihydro-1,4-benzothiazin-7-yl]thiophene-2-carboximidamide	IC50	28 nM	US-8618286: Benzoxazines, benzothiazines, and related compounds having NOS inhibitory activity
2-Ethyl-isothiourea	KI	29 nM
2-Propyl-isothiourea	KI	29 nM
CHEMBL4440481	KI	29 nM
CHEMBL5189279	KI	29 nM
(+/-)-N-{2-[2-(1-methyl-pyrrolidin-2-yl)-ethylamino]-benzothiazol-5-yl}-thiophene-2-carboxamidine	IC50	30 nM
CHEMBL4094689	KI	30 nM
CHEMBL4092027	KI	30 nM
(4R,7R)-4-Methyl-octahydro-[1]pyrindin-(2E)-ylideneamine	IC50	31 nM
CHEMBL4079543	KI	31 nM
6-[[(2S,4S)-4-[(6-amino-4-methyl-2-pyridinyl)methoxy]pyrrolidin-2-yl]methoxymethyl]-4-methylpyridin-2-amine	KI	31 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
CHEMBL4465991	KI	31 nM
CHEMBL4062164	KI	32 nM
6-[[(2R)-3-amino-2-[(6-amino-4-methyl-2-pyridinyl)methoxy]propoxy]methyl]-4-methylpyridin-2-amine	KI	32 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
CHEMBL4086610	KI	33 nM
N’-[4-[2-(2-hydroxyethylamino)ethyl]-2,3-dihydro-1,4-benzothiazin-7-yl]thiophene-2-carboximidamide	IC50	34 nM	US-8618286: Benzoxazines, benzothiazines, and related compounds having NOS inhibitory activity
6-[[(2R,4R)-4-[(6-amino-4-methyl-2-pyridinyl)methoxy]pyrrolidin-2-yl]methoxymethyl]-4-methylpyridin-2-amine	KI	34 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
N-{4-[2-(3-Chloro-benzylamino)-ethyl]-phenyl}-thiophene-2-carboxamidine; hydrochloride	IC50	35 nM
CHEMBL2333953	IC50	35 nM
(4S,7S)-4-Methyl-octahydro-[1]pyrindin-(2E)-ylideneamine	IC50	36 nM
CHEMBL4096049	KI	36 nM
6-[[(2S,3S)-2-amino-3-[(6-amino-4-methyl-2-pyridinyl)methoxy]butoxy]methyl]-4-methylpyridin-2-amine	KI	37 nM	US-9732037: 2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors
CHEMBL4450896	KI	37 nM
N-(4-Aminomethyl-phenyl)-acetamidine	KI	40 nM

ChEMBL bioactivities

435 potent at pChembl≥5 of 966 total, top 50 by pChembl (potency: 10 = 0.1 nM, 6 = 1 µM).

pChembl	Type	Value	Unit	Molecule
7.75	Ki	18	nM	CHEMBL105792
7.73	Ki	18.62	nM	CHEMBL1800346
7.66	Ki	22	nM	CHEMBL483091
7.62	Ki	24	nM	CHEMBL106871
7.52	Ki	30	nM	NITROARGININE
7.47	IC50	34	nM	CHEMBL302657
7.41	Ki	39	nM	ETHYLISOTHIOUREA
7.35	IC50	45	nM	CHEMBL59925
7.35	IC50	44.67	nM	CHEMBL59925
7.31	IC50	49	nM	CHEMBL294084
7.30	IC50	50	nM	CHEMBL8365
7.30	IC50	50	nM	CHEMBL302657
7.14	IC50	72	nM	CHEMBL40833
7.10	Ki	80	nM	CHEMBL227937
7.10	IC50	80	nM	CHEMBL1221702
7.10	IC50	80	nM	CHEMBL8292
7.09	IC50	81	nM	CHEMBL61427
7.07	Ki	85.2	nM	CHEMBL474637
7.02	Ki	95.2	nM	CHEMBL526688
7.00	IC50	100	nM	CHEMBL40833
6.97	Ki	107	nM	CHEMBL44833
6.96	Ki	110	nM	CHEMBL228077
6.92	IC50	120	nM	CHEMBL8281
6.84	IC50	146	nM	CHEMBL6808
6.82	IC50	150	nM	CHEMBL294609
6.82	IC50	150	nM	CHEMBL61939
6.80	IC50	160	nM	CHEMBL7945
6.77	IC50	170	nM	CHEMBL59925
6.75	IC50	180	nM	CHEMBL272708
6.75	IC50	180	nM	CHEMBL354788
6.70	IC50	200	nM	CHEMBL1230023
6.70	IC50	200	nM	CHEMBL299677
6.70	IC50	200	nM	CHEMBL1221703
6.66	IC50	220	nM	CHEMBL428069
6.66	IC50	220	nM	CHEMBL6808
6.64	IC50	230	nM	CHEMBL62077
6.64	IC50	230	nM	CHEMBL75304
6.62	IC50	240	nM	CHEMBL74919
6.60	Ki	250	nM	CHEMBL298404
6.60	IC50	250	nM	CHEMBL1783094
6.57	Ki	270	nM	CHEMBL49881
6.55	IC50	280	nM	CHEMBL268521
6.52	IC50	300	nM	TILARGININE
6.52	IC50	300	nM	CHEMBL40833
6.52	IC50	300	nM	CHEMBL269076
6.50	IC50	320	nM	NITROARGININE
6.50	IC50	320	nM	CHEMBL40833
6.50	IC50	320	nM	CHEMBL75484
6.50	Ki	320	nM	CHEMBL555794
6.48	IC50	330	nM	TILARGININE

PubChem BioAssay actives

393 with measured affinity, of 1385 total; 50 most potent distinct compounds. Largely complementary to BindingDB; screening values are coarse (µM, 4 dp), so sub-nM hits tie at the floor.

Compound	Assay	Type	Value	Unit
(1S,5S,6R,7R)-7-chloro-5-methyl-2-azabicyclo[4.1.0]hept-2-en-3-amine	2007174: Inhibition of human eNOS overexpressed in baculovirus infected Sf21 cells assessed as inhibition constant	ki	0.0180	uM
(1S,5S,6R,7R)-7-chloro-5-methyl-2-azabicyclo[4.1.0]hept-2-en-3-amine;hydrochloride	607321: Inhibition of human eNOS assessed as inhibition of [3H]L-arginine to [3H]L-citrulline conversion by scintillation counting	ki	0.0186	uM
propan-2-yl carbamimidothioate	2007166: Inhibition of eNOS in human DLD-1 cells assessed as reduction of L-[14C]arginine to L-[14C]citrulline conversion	ki	0.0220	uM
(2S)-2-amino-5-[[amino(ethylsulfanyl)methylidene]amino]pentanoic acid	2007149: Inhibition of human eNOS assessed as inhibition constant	ki	0.0240	uM
(2S)-2-amino-5-[[amino(nitramido)methylidene]amino]pentanoic acid	68156: Inhibitory activity against human endothelial nitric oxide synthase (eNOS) isoenzyme.	ki	0.0300	uM
4-methyl-6-propylpyridin-2-amine	1799391: NOS Enzyme Inhibition Assay from Article 10.1038/nchembio.115: “Anchored plasticity opens doors for selective inhibitor design in nitric oxide synthase.”	ic50	0.0340	uM
ethyl carbamimidothioate	2007166: Inhibition of eNOS in human DLD-1 cells assessed as reduction of L-[14C]arginine to L-[14C]citrulline conversion	ki	0.0390	uM
4,6-dimethylpyridin-2-amine	607321: Inhibition of human eNOS assessed as inhibition of [3H]L-arginine to [3H]L-citrulline conversion by scintillation counting	ic50	0.0447	uM
6-ethyl-4-methylpyridin-2-amine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.0490	uM
5-cyclopropyl-4,5-dihydrothieno[2,3-c]pyridin-7-amine	85462: Ability to inhibit the conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by e-NOS from HUVEC cells	ic50	0.0500	uM
4-methylpyridin-2-amine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.0720	uM
5-ethynyl-4,5-dihydrothieno[2,3-c]pyridin-7-amine	85462: Ability to inhibit the conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by e-NOS from HUVEC cells	ic50	0.0800	uM
(2S)-2-methyl-2,3-dihydrothieno[2,3-f][1,4]oxazepin-5-amine	1799391: NOS Enzyme Inhibition Assay from Article 10.1038/nchembio.115: “Anchored plasticity opens doors for selective inhibitor design in nitric oxide synthase.”	ic50	0.0800	uM
2-[(4S)-4-amino-5-(2-aminoethylamino)pentyl]-1-nitroguanidine	414630: Inhibition of eNOS	ki	0.0800	uM
4-methylpyridine-2,3-diamine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.0810	uM
N’-[(3S,4S)-4-[(6-amino-4-methyl-2-pyridinyl)methyl]pyrrolidin-3-yl]-N-[(4-chlorophenyl)methyl]ethane-1,2-diamine	414630: Inhibition of eNOS	ki	0.0852	uM
N’-[(3R,4S)-4-[(6-amino-4-methyl-2-pyridinyl)methyl]pyrrolidin-3-yl]-N-[(3-chlorophenyl)methyl]ethane-1,2-diamine	414630: Inhibition of eNOS	ki	0.0952	uM
(2S)-2-amino-5-[[amino(nitramido)methylidene]amino]-N-[(2S)-1,4-diamino-1-oxobutan-2-yl]pentanamide	414630: Inhibition of eNOS	ki	0.1070	uM
(2S,4R)-4-[[(2S)-2-amino-5-[[amino(nitramido)methylidene]amino]pentanoyl]amino]pyrrolidine-2-carboxamide	414630: Inhibition of eNOS	ki	0.1100	uM
6-ethynyl-6,7-dihydrothieno[3,2-c]pyridin-4-amine	85462: Ability to inhibit the conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by e-NOS from HUVEC cells	ic50	0.1200	uM
4-methyl-2,3,4,5-tetrahydropyridin-6-amine	241405: Inhibitory activity against human Endothelial nitric oxide synthase (eNOS)	ic50	0.1460	uM
4-methyl-6-(2-methylpropyl)pyridin-2-amine	415183: Inhibition of human eNOS	ic50	0.1500	uM
3,4-dimethylpyridin-2-amine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.1500	uM
8-fluoro-3-(4-fluorophenyl)-3,4-dihydroisoquinolin-1-amine	85322: Ability to inhibit conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by endothelial NOS (e NOS) from HUVEC cells	ic50	0.1600	uM
5,8-difluoro-2-(furan-2-yl)-1,2-dihydroquinazolin-4-amine	68135: Inhibitory activity against human endothelial nitric oxide synthase	ic50	0.1800	uM
N-[(5-chloro-2-nitrophenyl)methyl]-N,1-dimethylpiperidin-4-amine	316793: Inhibition of human eNOS expressed in insect SF9 cells after 1 hr	ic50	0.1800	uM
(3R)-3-propyl-2,3-dihydrothieno[2,3-f][1,4]oxazepin-5-amine	1799391: NOS Enzyme Inhibition Assay from Article 10.1038/nchembio.115: “Anchored plasticity opens doors for selective inhibitor design in nitric oxide synthase.”	ic50	0.2000	uM
(3S)-3-propyl-2,3-dihydrothieno[2,3-f][1,4]oxazepin-5-amine	693738: Inhibition of wild type human eNOS using L-Arg as substrate incubated for 1 hr prior to L-Arg addition	ic50	0.2000	uM
4-methyl-6-propan-2-ylpyridin-2-amine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.2000	uM
4-chloro-2-[[methyl-[1-(2-phenylethyl)piperidin-4-yl]amino]methyl]benzonitrile	316793: Inhibition of human eNOS expressed in insect SF9 cells after 1 hr	ic50	0.2200	uM
4-ethylpyridin-2-amine	68131: Inhibitory activity against endothelial nitric oxide synthase (eNOS)	ic50	0.2300	uM
4-methyl-2,3-dihydropyridin-6-amine	67822: Inhibition of human endothelial Nitric Oxide Synthase expressed in Sf-21 cells	ic50	0.2300	uM
2,4-dimethyl-2,3-dihydropyridin-6-amine	67822: Inhibition of human endothelial Nitric Oxide Synthase expressed in Sf-21 cells	ic50	0.2400	uM
(2R)-azetidine-2-carboximidamide	600731: Inhibition of human recombinant eNOS by microtiter plate assay	ic50	0.2500	uM
4,5-dihydrothieno[2,3-c]pyridin-7-amine	85462: Ability to inhibit the conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by e-NOS from HUVEC cells	ic50	0.2800	uM
(2S)-2-amino-5-[(N’-methylcarbamimidoyl)amino]pentanoic acid	85322: Ability to inhibit conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by endothelial NOS (e NOS) from HUVEC cells	ic50	0.3000	uM
2,4-dimethyl-2,3,4,5-tetrahydropyridin-6-amine	68142: inhibition of human endothelial constitutive Endothelial nitric oxide synthase (heNOS)	ic50	0.3000	uM
4,5-dimethyl-2,3-dihydropyridin-6-amine	67822: Inhibition of human endothelial Nitric Oxide Synthase expressed in Sf-21 cells	ic50	0.3200	uM
N’-[3-(aminomethyl)phenyl]-2-methylsulfanylethanimidamide;dihydrobromide	68156: Inhibitory activity against human endothelial nitric oxide synthase (eNOS) isoenzyme.	ki	0.3200	uM
N’-[3-(aminomethyl)phenyl]furan-2-carboximidamide;dihydrobromide	68156: Inhibitory activity against human endothelial nitric oxide synthase (eNOS) isoenzyme.	ki	0.3500	uM
N’-[3-(aminomethyl)phenyl]furan-2-carboximidamide	2007183: Inhibition of human eNOS assessed as reduction of L-[14C]arginine to L-[14C]-citrulline conversion	ki	0.3500	uM
(4R,4aR,7aR)-4-methyl-4,4a,5,6,7,7a-hexahydro-3H-cyclopenta[b]pyridin-2-amine	241405: Inhibitory activity against human Endothelial nitric oxide synthase (eNOS)	ic50	0.3600	uM
4-methyl-2-prop-2-enyl-2,3-dihydropyridin-6-amine	67822: Inhibition of human endothelial Nitric Oxide Synthase expressed in Sf-21 cells	ic50	0.3600	uM
3-[[(3S,4S)-4-[(6-amino-2-pyridinyl)methyl]pyrrolidin-3-yl]amino]propan-1-ol	414630: Inhibition of eNOS	ki	0.3666	uM
6-[4-[2-[4-(2-methylpropyl)piperazin-1-yl]ethyl]phenyl]pyridin-2-amine	68133: Inhibitory activity against human endothelial nitric oxide synthase (eNOS)	ic50	0.3720	uM
(2S)-2-amino-5-[[amino(methylsulfanyl)methylidene]amino]pentanoic acid	241624: Inhibitory concentration against human endothelial nitric oxide synthase expressed in Sf-9 cells	ic50	0.4000	uM
ethyl N’-phenylcarbamimidothioate;hydroiodide	68155: Inhibitory activity against human vascular endothelial nitric oxide synthase.	ki	0.4000	uM
ethyl N’-phenylcarbamimidothioate	2007172: Inhibition of human eNOS assessed as reduction of L-[14C]arginine to L-[14C]citrulline conversion	ki	0.4000	uM
N’-[3-(aminomethyl)phenyl]thiophene-2-carboximidamide;dihydrobromide	68156: Inhibitory activity against human endothelial nitric oxide synthase (eNOS) isoenzyme.	ki	0.4000	uM
N’-[1-(2-pyrrolidin-1-ylethyl)-2,3-dihydroindol-5-yl]thiophene-2-carboximidamide;dihydrochloride	715215: Inhibition of recombinant human iNOS expressed in baculovirus-infected Sf9 cells assessed as conversion of [3H]-larginine to [3H]-L-citrulline preincubated for 15 mins measured after 45 mins by liquid scintillation counting	ic50	0.4100	uM

CTD chemical–gene interactions

285 total (human), top 30 by PubMed support.

Chemical	Actions (top 5)	PubMed papers
Resveratrol	decreases reaction, increases phosphorylation, affects reaction, decreases activity, increases activity (+11 more)	28
Nitric Oxide	increases abundance, decreases expression, decreases activity, increases expression, increases activity (+10 more)	21
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one	increases abundance, affects cotreatment, decreases phosphorylation, decreases reaction, increases phosphorylation (+5 more)	13
Glucose	affects cotreatment, affects reaction, decreases phosphorylation, decreases reaction, decreases expression (+4 more)	11
bisphenol A	affects cotreatment, decreases methylation, decreases expression, increases expression, increases activity (+2 more)	10
Hydrogen Peroxide	decreases chemical synthesis, decreases secretion, decreases expression, decreases reaction, increases expression (+5 more)	10
Wortmannin	increases activity, increases phosphorylation, affects cotreatment, increases expression, increases reaction (+3 more)	8
Simvastatin	affects cotreatment, decreases reaction, increases expression, increases phosphorylation, increases reaction (+2 more)	8
U 0126	decreases expression, decreases reaction, increases phosphorylation, increases activity, increases abundance (+1 more)	6
Fulvestrant	decreases methylation, decreases reaction, increases expression, increases phosphorylation, increases abundance (+1 more)	6
Estradiol	increases phosphorylation, increases chemical synthesis, decreases reaction, increases activity, increases reaction (+2 more)	6
Plant Extracts	decreases expression, decreases reaction, increases expression, increases activity, increases phosphorylation (+1 more)	6
Quercetin	decreases reaction, increases phosphorylation, increases expression, decreases expression	6
1,2-bis(2-aminophenoxy)ethane N,N,N’,N’-tetraacetic acid acetoxymethyl ester	increases phosphorylation, increases abundance, decreases reaction, increases expression	4
dorsomorphin	decreases phosphorylation, decreases reaction, increases activity, increases phosphorylation, increases expression	4
Atorvastatin	affects response to substance, decreases expression, decreases reaction, increases expression, increases reaction	4
Acetylcysteine	decreases reaction, increases expression, decreases expression	4
Calcium	decreases reaction, increases phosphorylation, affects reaction, increases activity, increases expression	4
Oxygen	decreases reaction, increases expression, decreases expression, decreases activity	4
Tobacco Smoke Pollution	increases expression, decreases phosphorylation, decreases expression	4
Genistein	decreases reaction, increases expression, decreases expression, increases activity	4
Rosiglitazone	decreases phosphorylation, decreases reaction, increases abundance, increases phosphorylation	3
Arsenic Trioxide	decreases expression, increases expression, increases activity, increases phosphorylation	3
Air Pollutants	increases abundance, increases expression, affects response to substance, decreases methylation	3
Benzo(a)pyrene	decreases reaction, affects methylation, increases methylation, increases phosphorylation	3
Dibutyl Phthalate	decreases phosphorylation, increases expression, increases phosphorylation, decreases reaction	3
Folic Acid	decreases reaction, decreases activity, affects cotreatment, decreases expression, affects phosphorylation	3
Methotrexate	decreases expression, increases expression, affects response to substance	3
Progesterone	increases expression, increases phosphorylation, decreases expression, decreases reaction, increases activity (+1 more)	3
Superoxides	affects chemical synthesis, decreases reaction, increases abundance, increases reaction, increases activity (+2 more)	3

ChEMBL screening assays

188 unique, capped per target: 178 binding, 6 admet, 4 functional

Representative assays (with source publication via chembl_document):

Assay ID	Type	Description	Source paper
CHEMBL675570	Binding	Ratio of Endothelial Nitric Oxide Synthase to Inducible nitric oxide synthase	Inhibition of inducible nitric oxide synthase by acetamidine derivatives of hetero-substituted lysine and homolysine. — *Bioorg Med Chem Lett*
CHEMBL4032165	ADMET	Inhibition of human endothelial NOS expressed in Escherichia coli using L-arginine as substrate assessed as reduction in NO production by measuring oxidation of oxyHb to metHb measured for 6 mins by hemoglobin- NO capture assay	Improvement of Cell Permeability of Human Neuronal Nitric Oxide Synthase Inhibitors Using Potent and Selective 2-Aminopyridine-Based Scaffolds with a Fluorobenzene Linker. — *J Med Chem*
CHEMBL690699	Functional	Ability to inhibit conversion of [3H]L-Arg to [3H]L-citrulline catalyzed by endothelial NOS (e NOS) from HUVEC cells	3,4-Dihydro-1-isoquinolinamines: a novel class of nitric oxide synthase inhibitors with a range of isoform selectivity and potency. — *Bioorg Med Chem Lett*

Cellosaurus cell lines

2 cell lines: 2 cancer cell line

First 10 cell lines (id-ordered, not curated):

Cellosaurus	Name	Category	Sex
CVCL_D8RI	Ubigene HCT 116 NOS3 KO	Cancer cell line	Male
CVCL_F6HH	EDITGENE NOS3 Knockout A-549 Cell Line	Cancer cell line	Male

Clinical trials (associated diseases)

Via MONDO — disease-level, not drug-specific.

showing 60 of 300 trials:

NCT	Phase	Status	Title
NCT00009191	Phase 4	COMPLETED	The Depression in Alzheimer’s Disease Study (DIADS)
NCT00009217	Phase 4	COMPLETED	Treatment of Behavioral Symptoms in Alzheimer’s Disease
NCT00018278	Phase 4	COMPLETED	Electrophysiologic Measures of Treatment Response in Alzheimer Disease
NCT00035204	Phase 4	COMPLETED	A Study of the Effects on Sleep, Attention, and Gastrointestinal Tolerance of Galantamine and Donepezil in Patients With Alzheimer’s Disease
NCT00042172	Phase 4	COMPLETED	Treatment for Early Memory Loss
NCT00046358	Phase 4	COMPLETED	The Effect of Short-Term Statins and NSAIDs on Levels of Beta-Amyloid, a Protein Associated With Alzheimer’s Disease
NCT00104442	Phase 4	COMPLETED	Study of the Effects of Current Drug Treatments on Levels of Certain Brain Chemicals in Alzheimer’s Disease
NCT00120874	Phase 4	COMPLETED	Memantine and Comprehensive, Individualized Management of Alzheimer’s Disease and Caregiver Training
NCT00142324	Phase 4	UNKNOWN	CALM-AD
NCT00165724	Phase 4	COMPLETED	Alzheimer’s Disease Long-term Follow-up Study (ALF Study)
NCT00165750	Phase 4	TERMINATED	Correlation Between Regional Brain Volume and Response to Donepezil Treatment in AD Patients
NCT00202124	Phase 4	COMPLETED	Double Blind Study of Trp01 in Patients With Alzheimer’s Disease
NCT00208819	Phase 4	COMPLETED	A Comparison of Two Standard Therapies in the Management of Dementia With Agitation
NCT00216515	Phase 4	COMPLETED	The Efficacy of Galantamine on the Attention and the Frontal Function of the Patients With Dementia of Alzheimer Type
NCT00230568	Phase 4	COMPLETED	EARTH 413: A Study of Aricept in Hispanic Patients With Mild to Moderate Alzheimer’s Disease (AD)
NCT00234637	Phase 4	COMPLETED	Rivastigmine Monotherapy and Combination Therapy With Memantine in Patients With Moderately Severe Alzheimer’s Disease Who Failed to Benefit From Previous Cholinesterase Inhibitor Treatment
NCT00245206	Phase 4	COMPLETED	Side Effects of Newer Antipsychotics in Older Adults
NCT00254033	Phase 4	COMPLETED	Apathy Associated With Alzheimer’s Disease
NCT00260624	Phase 4	COMPLETED	Escitalopram Treatment of Patients With Agitated Dementia
NCT00303277	Phase 4	COMPLETED	Do HMG CoA Reductase Inhibitors Affect Abeta Levels?
NCT00305903	Phase 4	COMPLETED	Safety and Tolerability of Rivastigmine With Add-on Memantine in Patients With Probable Alzheimer’s Disease
NCT00306124	Phase 4	UNKNOWN	Dopaminergic Enhancement of Learning and Memory in Healthy Adults and Patients With Dementia/Mild Cognitive Impairment
NCT00334906	Phase 4	COMPLETED	Study of Memantine in Assessment of Selected Measures of Volumetric Magnetic Resonance Imaging (MRI) and Cognition in Moderate AD (Alzheimer’s Disease)
NCT00369603	Phase 4	TERMINATED	Functional Brain Imaging of Medication Treatment Response in Mild Alzheimer’s Disease Patients
NCT00375557	Phase 4	WITHDRAWN	Safety and Efficacy of Divalproex and Quetiapine in Elderly Alzheimer’s Dementia Patients
NCT00381381	Phase 4	COMPLETED	The Clinical Response of Choline Acetyltransferase and Apolipoprotein Epsilon Gene Polymorphisms to Donepezil in Alzheimer’s Disease
NCT00385684	Phase 4	COMPLETED	Low-Dose Opiate Therapy for Discomfort in Dementia (L-DOT)
NCT00401167	Phase 4	COMPLETED	Memantine for Agitation and Aggression in Severe Alzheimer’s Disease
NCT00403520	Phase 4	COMPLETED	Hippocampus Study: Comparative Effect of Donepezil 10mg/d and Placebo on Clinical and Radiological Markers
NCT00417482	Phase 4	COMPLETED	Antipsychotic Discontinuation in Alzheimer’s Disease
NCT00443014	Phase 4	COMPLETED	The Dementia Study in Northern Norway
NCT00469456	Phase 4	COMPLETED	Effect of Memantine on Functional Communication in Patients With Alzheimer’s Disease
NCT00476008	Phase 4	COMPLETED	Delaying the Progression of Driving Impairment in Individuals With Mild Alzheimer’s Disease
NCT00477659	Phase 4	COMPLETED	Neural Correlates In Mild Alzheimer’s Disease
NCT00480870	Phase 4	COMPLETED	The Effect of Anticholinesterase Drugs on Sleep in Alzheimer’s Disease Patients
NCT00495820	Phase 4	COMPLETED	Methylphenidate for Apathy in Alzheimer’s Dementia: A Controlled Study
NCT00523666	Phase 4	UNKNOWN	Diffusion Tensor Weighted MRI in Alzheimer’s Disease Modifying Treatment Effects of Galantamine (Reminyl®)
NCT00549601	Phase 4	COMPLETED	Convenience, Tolerability, and Safety of Change in the Administration of Rivastigmine From Capsules to a Transdermal Patch in Patients With Mild to Moderate Alzheimer’s Disease
NCT00551161	Phase 4	COMPLETED	Magnetic Resonance Spectroscopy Study of Memantine in Alzheimer’s Disease
NCT00561392	Phase 4	COMPLETED	Clinical Effectiveness of 10 cm^2 Rivastigmine Patch in Patients With Alzheimer’s Disease
NCT00622713	Phase 4	COMPLETED	A 24 Week, Multicenter, Open, Evaluation of the Clinical Effectiveness of the Once-daily 10 cm^2 Rivastigmine Patch Formulation in Patients With Probable Alzheimer’s Disease (EXTRA)
NCT00626210	Phase 4	TERMINATED	Modafinil Treatment for Sleep/Wake Disturbances in Older Adults
NCT00626613	Phase 4	UNKNOWN	The Relationship Between Risperdal Treatment and Quality of Life in Patients With Alzheimer’s Disease and Behavioural and Psychological Symptoms of Dementia (BPSD)
NCT00649220	Phase 4	TERMINATED	Memantine and Antipsychotics Use
NCT00706186	Phase 4	TERMINATED	Safety and Feasibility of Sodium Oxybate in Mild Alzheimer’s Disease Patients
NCT00711204	Phase 4	WITHDRAWN	Evaluating The Impact Of Donepezil Hydrochloride (Aricept) On Behavioral And Psychological Symptoms In Patients With Severe Alzheimer’s Disease
NCT00731224	Phase 4	COMPLETED	Compliance and Tolerability of Rivastigmine Transdermal Patch 10 cm² in Patients With Probable Alzheimer’s Disease.
NCT00792662	Phase 4	WITHDRAWN	Improving Function, Quality of Life, Glycemia in Diabetics With Dementia
NCT00800709	Phase 4	COMPLETED	Memantine and Changes of Biological Markers and Brain PET Imaging in Alzheimer’s Disease
NCT00862940	Phase 4	COMPLETED	A Clinical Study Evaluating the Effects of Memantine on Brain Atrophy in Patients With Alzheimer’s Disease
NCT00933608	Phase 4	COMPLETED	Effects of Memantine on Magnetic Resonance (MR) Spectroscopy in Subjects at Risk for Alzheimer’s Disease
NCT00948766	Phase 4	COMPLETED	Effects of Rivastigmine Patch on Activities of Daily Living and Cognition in Patients With Severe Dementia of the Alzheimer’s Type (ACTION) (Study Protocol CENA713DUS44, NCT00948766) and a 24 Week Open-label Extension to Study CENA713DUS44
NCT00950430	Phase 4	ENROLLING_BY_INVITATION	Imaging of Brain Amyloid Plaques in the Aging Population
NCT00980785	Phase 4	COMPLETED	Studying the Effects of Antihypertensives on Individuals at Risk for Alzheimer’s
NCT01025466	Phase 4	COMPLETED	Exelon Patch and Combination With Memantine Comparative Trial
NCT01047579	Phase 4	COMPLETED	A 12 Week, Multicenter, Open Label Evaluation of Caregiver Preference, Safety and Tolerability of Exelon® Patch (Rivastigmine Transdermal) in Patients With Alzheimer’s Disease
NCT01054976	Phase 4	COMPLETED	The Efficacy of Galantamine Treatment on Attention in Patients With Alzheimer’s Disease
NCT01119638	Phase 4	COMPLETED	Escitalopram Treatment for BPSD in Alzheimer’s Disease in Comparison to Risperidone
NCT01122329	Phase 4	COMPLETED	A Positron Emission Tomography (PET) Study Evaluating Brain Metabolism of a Medical Food in Alzheimer’s Disease
NCT01142336	Phase 4	COMPLETED	Effects of Simvastatin on Biomarkers

Predicted chemistry — Sugi Predict

The predicted patent-compound landscape for NOS3 across ~30M SureChEMBL compounds — explore on Sugi Predict.