Reviewed, UniProtKB/Swiss-Prot P29473 (NOS3_BOVIN)

Names and origin

Protein names

Recommended name:
    Nitric oxide synthase, endothelial
    EC=1.14.13.39
Alternative name(s):
    Endothelial NOS
      Short name=eNOS
    EC-NOS
    NOS type III
      Short name=NOSIII
    Constitutive NOS
      Short name=cNOS

Gene names

Name:

NOS3

Organism

Bos taurus (Bovine)

Taxonomic identifier

9913 [NCBI]

Taxonomic lineage

Eukaryota › Metazoa › Chordata › Craniata › Vertebrata › Euteleostomi › Mammalia › Eutheria › Laurasiatheria › Cetartiodactyla › Ruminantia › Pecora › Bovidae › Bovinae › Bos

Protein attributes

Sequence length	1205 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level.

General annotation (Comments)

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.
Catalytic activity	L-arginine + n NADPH + n H+ + m O2 = citrulline + nitric oxide + n NADP+.
Cofactor	Heme group. Binds 1 FAD. Binds 1 FMN. Tetrahydrobiopterin (BH4). May stabilize the dimeric form of the enzyme.
Enzyme regulation	Stimulated by calcium/calmodulin. Inhibited by NOSIP and NOSTRIN By similarity.
Subunit structure	Homodimer. Interacts with NOSIP and NOSTRIN By similarity.
Subcellular location	Cell membrane. Membrane › caveola. Cytoplasm › cytoskeleton By similarity. Golgi apparatus. Note: Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle; which is favored by interaction with NOSIP and results in a reduced enzymatic activity By similarity. Ref.7
Sequence similarities	Belongs to the NOS family. Contains 1 FAD-binding FR-type domain. Contains 1 flavodoxin-like domain.

Ontologies

Keywords
Biological process	Blood coagulation
Cellular component	Cell membrane Cytoplasm Cytoskeleton Golgi apparatus Membrane
Ligand	Calcium Calmodulin-binding FAD FMN Heme Iron Metal-binding NADP Zinc
Molecular function	Oxidoreductase
PTM	Lipoprotein Myristate Palmitate Phosphoprotein
Technical term	3D-structure
Gene Ontology (GO)
Biological process	anti-apoptosis Inferred from mutant phenotype. Source: UniProtKB arginine catabolic process Inferred from direct assay. Source: UniProtKB blood coagulation Inferred from electronic annotation. Source: UniProtKB-KW mitochondrion organization Inferred from mutant phenotype. Source: UniProtKB nitric oxide biosynthetic process Inferred from direct assay. Source: UniProtKB oxidation reduction Inferred from electronic annotation. Source: UniProtKB-KW positive regulation of guanylate cyclase activity Inferred from mutant phenotype. Source: UniProtKB
Cellular component	Golgi apparatus Inferred from direct assay. Source: UniProtKB caveola Inferred from electronic annotation. Source: UniProtKB-SubCell cytoskeleton Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	FAD binding Inferred from electronic annotation. Source: InterPro FMN binding Inferred from electronic annotation. Source: InterPro NADP or NADPH binding Inferred from electronic annotation. Source: InterPro calcium ion binding Inferred from electronic annotation. Source: UniProtKB-KW calmodulin binding Inferred from electronic annotation. Source: UniProtKB-KW heme binding Inferred from electronic annotation. Source: InterPro nitric-oxide synthase activity Inferred from direct assay. Source: UniProtKB zinc ion binding Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed

Chain

2 – 1205

1204

Nitric oxide synthase, endothelial

PRO_0000170941

Regions

Domain

522 – 705

184

Flavodoxin-like

Domain

758 – 1004

247

FAD-binding FR-type

Nucleotide binding

651 – 682

32

FMN By similarity

Nucleotide binding

795 – 806

12

FAD By similarity

Nucleotide binding

937 – 947

11

FAD By similarity

Nucleotide binding

1012 – 1030

19

NADP By similarity

Nucleotide binding

1110 – 1125

16

NADP By similarity

Region

100 – 488

389

Interaction with NOSIP By similarity

Region

492 – 512

21

Calmodulin-binding Potential

Sites

Metal binding

96

1

Zinc

Metal binding

101

1

Zinc

Metal binding

186

1

Iron (heme axial ligand)

Amino acid modifications

Modified residue

83

1

Phosphotyrosine By similarity

Modified residue

143

1

Phosphoserine; by PKA

Modified residue

497

1

Phosphothreonine; by PKA Ref.6

Modified residue

635

1

Phosphoserine Ref.6

Modified residue

1177

1

Phosphothreonine By similarity

Modified residue

1179

1

Phosphoserine; by PDPK1 and PKA Ref.6

Lipidation

2

1

N-myristoyl glycine Ref.4

Lipidation

15

1

S-palmitoyl cysteine Ref.5

Lipidation

26

1

S-palmitoyl cysteine Ref.5

Experimental info

Sequence conflict

100

1

C → R in AAA30669. Ref.3

Sequence conflict

165

1

Y → I in AAA30669. Ref.3

Sequence conflict

318 – 328

11

EHPTLEWFAAL → GAPHTGVVRGP in AAA30669. Ref.3

Sequence conflict

455

1

S → Y in AAA30669. Ref.3

Sequence conflict

459

1

T → P in AAA30669. Ref.3

Sequence conflict

741

1

T → A in AAA30669. Ref.3

Sequence conflict

804 – 805

2

CP → SA in AAA30669. Ref.3

Sequence conflict

857

1

L → V in AAA30669. Ref.3

Sequence conflict

907 – 908

2

WF → LV in AAA30669. Ref.3

Sequence conflict

1042

1

A → H in AAA30669. Ref.3

Secondary structure

1

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1205

Helix Strand Turn

Details...

Beta strand

72 – 75

4

Turn

76 – 79

4

Beta strand

80 – 83

4

Helix

86 – 89

4

Turn

109 – 111

3

Helix

122 – 139

18

Helix

146 – 162

17

Helix

169 – 181

13

Helix

189 – 191

3

Beta strand

196 – 199

4

Helix

206 – 221

16

Helix

222 – 224

3

Beta strand

229 – 232

4

Beta strand

247 – 251

5

Beta strand

255 – 257

3

Beta strand

263 – 265

3

Helix

267 – 269

3

Helix

270 – 278

9

Beta strand

286 – 288

3

Beta strand

293 – 296

4

Beta strand

303 – 305

3

Helix

309 – 311

3

Beta strand

314 – 316

3

Helix

323 – 328

6

Beta strand

331 – 334

4

Beta strand

342 – 345

4

Beta strand

348 – 351

4

Helix

361 – 365

5

Helix

367 – 370

4

Turn

372 – 375

4

Helix

378 – 384

7

Helix

392 – 394

3

Helix

396 – 414

19

Helix

422 – 440

19

Helix

447 – 450

4

Beta strand

453 – 455

3

Helix

456 – 458

3

Helix

460 – 463

4

Beta strand

472 – 476

5

Sequences

	Sequence		Length	Mass (Da)
	P29473-1 [UniParc]. Last modified January 23, 2007. Version 3. Checksum: 5DC8FF4F25870281	FASTA	1,205	133,287
	10 20 30 40 50 60 MGNLKSVGQE PGPPCGLGLG LGLGLCGKQG PASPAPEPSR APAPATPHAP DHSPAPNSPT 70 80 90 100 110 120 LTRPPEGPKF PRVKNWELGS ITYDTLCAQS QQDGPCTPRC CLGSLVLPRK LQTRPSPGPP 130 140 150 160 170 180 PAEQLLSQAR DFINQYYSSI KRSGSQAHEE RLQEVEAEVA STGTYHLRES ELVFGAKQAW 190 200 210 220 230 240 RNAPRCVGRI QWGKLQVFDA RDCSSAQEMF TYICNHIKYA TNRGNLRSAI TVFPQRAPGR 250 260 270 280 290 300 GDFRIWNSQL VRYAGYRQQD GSVRGDPANV EITELCIQHG WTPGNGRFDV LPLLLQAPDE 310 320 330 340 350 360 APELFVLPPE LVLEVPLEHP TLEWFAALGL RWYALPAVSN MLLEIGGLEF SAAPFSGWYM 370 380 390 400 410 420 STEIGTRNLC DPHRYNILED VAVCMDLDTR TTSSLWKDKA AVEINLAVLH SFQLAKVTIV 430 440 450 460 470 480 DHHAATVSFM KHLDNEQKAR GGCPADWAWI VPPISGSLTP VFHQEMVNYI LSPAFRYQPD 490 500 510 520 530 540 PWKGSATKGA GITRKKTFKE VANAVKISAS LMGTLMAKRV KATILYASET GRAQSYAQQL 550 560 570 580 590 600 GRLFRKAFDP RVLCMDEYDV VSLEHEALVL VVTSTFGNGD PPENGESFAA ALMEMSGPYN 610 620 630 640 650 660 SSPRPEQHKS YKIRFNSVSC SDPLVSSWRR KRKESSNTDS AGALGTLRFC VFGLGSRAYP 670 680 690 700 710 720 HFCAFARAVD TRLEELGGER LLQLGQGDEL CGQEEAFRGW AKAAFQASCE TFCVGEEAKA 730 740 750 760 770 780 AAQDIFSPKR SWKRQRYRLS TQAEGLQLLP GLIHVHRRKM FQATVLSVEN LQSSKSTRAT 790 800 810 820 830 840 ILVRLDTAGQ EGLQYQPGDH IGICPPNRPG LVEALLSRVE DPPPPTESVA VEQLEKGSPG 850 860 870 880 890 900 GPPPSWVRDP RLPPCTLRQA LTFFLDITSP PSPRLLRLLS TLAEEPSEQQ ELETLSQDPR 910 920 930 940 950 960 RYEEWKWFRC PTLLEVLEQF PSVALPAPLL LTQLPLLQPR YYSVSSAPNA HPGEVHLTVA 970 980 990 1000 1010 1020 VLAYRTQDGL GPLHYGVCST WLSQLKTGDP VPCFIRGAPS FRLPPDPYVP CILVGPGTGI 1030 1040 1050 1060 1070 1080 APFRGFWQER LHDIESKGLQ PAPMTLVFGC RCSQLDHLYR DEVQDAQERG VFGRVLTAFS 1090 1100 1110 1120 1130 1140 REPDSPKTYV QDILRTELAA EVHRVLCLER GHMFVCGDVT MATSVLQTVQ RILATEGDME 1150 1160 1170 1180 1190 1200 LDEAGDVIGV LRDQQRYHED IFGLTLRTQE VTSRIRTQSF SLQERHLRGA VPWAFDPPGP DTPGP

References

[1]	"Endothelial nitric oxide synthase: molecular cloning and characterization of a distinct constitutive enzyme isoform." Lamas S., Marsden P.A., Li G.K., Tempst P., Michel T. Proc. Natl. Acad. Sci. U.S.A. 89:6348-6352(1992) [PubMed: 1378626] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[2]	"Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase." Nishida K., Harrison D.G., Navas J.P., Fisher A.A., Dockery S.P., Uematsu M., Nerem R.M., Alexander R.W., Murphy T.J. J. Clin. Invest. 90:2092-2096(1992) [PubMed: 1385480] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[3]	"Molecular cloning and expression of a cDNA encoding endothelial cell nitric oxide synthase." Sessa W.C., Harrison J.K., Barber C.M., Zeng D., Durieux M.E., D'Angelo D.D., Lynch K.R., Peach M.J. J. Biol. Chem. 267:15274-15276(1992) [PubMed: 1379225] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA]. Tissue: Aortic endothelium.
[4]	"Endothelial nitric oxide synthase. N-terminal myristoylation determines subcellular localization." Busconi L., Michel T. J. Biol. Chem. 268:8410-8413(1993) [PubMed: 7682550] [Abstract] Cited for: MYRISTOYLATION AT GLY-2.
[5]	"Mutagenesis of palmitoylation sites in endothelial nitric oxide synthase identifies a novel motif for dual acylation and subcellular targeting." Robinson L.J., Michel T. Proc. Natl. Acad. Sci. U.S.A. 92:11776-11780(1995) [PubMed: 8524847] [Abstract] Cited for: PALMITOYLATION AT CYS-15 AND CYS-26.
[6]	"Shear stress stimulates phosphorylation of eNOS at Ser(635) by a protein kinase A-dependent mechanism." Boo Y.C., Hwang J., Sykes M., Michell B.J., Kemp B.E., Lum H., Jo H. Am. J. Physiol. 283:H1819-H1828(2002) [PubMed: 12384459] [Abstract] Cited for: PHOSPHORYLATION AT THR-497; SER-635 AND SER-1179.
[7]	"Aberrant cytoplasmic sequestration of eNOS in endothelial cells after monocrotaline, hypoxia, and senescence: live-cell caveolar and cytoplasmic NO imaging." Mukhopadhyay S., Xu F., Sehgal P.B. Am. J. Physiol. 292:H1373-H1389(2007) [PubMed: 17071725] [Abstract] Cited for: SUBCELLULAR LOCATION.
[8]	"Crystal structure of constitutive endothelial nitric oxide synthase: a paradigm for pterin function involving a novel metal center." Raman C.S., Li H., Martasek P., Kral V., Masters B.S.S., Poulos T.L. Cell 95:939-950(1998) [PubMed: 9875848] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF 67-482.
[9]	"Mapping the active site polarity in structures of endothelial nitric oxide synthase heme domain complexed with isothioureas." Li H., Raman C.S., Martasek P., Kral V., Masters B.S.S., Poulos T.L. J. Inorg. Biochem. 81:133-139(2000) [PubMed: 11051558] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.86 ANGSTROMS) OF 67-482.
[10]	"Crystallographic studies on endothelial nitric oxide synthase complexed with nitric oxide and mechanism-based inhibitors." Li H., Raman C.S., Martasek P., Masters B.S.S., Poulos T.L. Biochemistry 40:5399-5406(2001) [PubMed: 11331003] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.93 ANGSTROMS).
[11]	"Crystal structure of nitric oxide synthase bound to nitro indazole reveals a novel inactivation mechanism." Raman C.S., Li H., Martasek P., Southan G., Masters B.S.S., Poulos T.L. Biochemistry 40:13448-13455(2001) [PubMed: 11695891] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS).
[12]	"Implications for isoform-selective inhibitor design derived from the binding mode of bulky isothioureas to the heme domain of endothelial nitric-oxide synthase." Raman C.S., Li H., Martasek P., Babu B.R., Griffith O.W., Southan G., Masters B.S.S., Poulos T.L. J. Biol. Chem. 276:26486-26491(2001) [PubMed: 11331290] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.93 ANGSTROMS).
[13]	"Structural basis for pterin antagonism in nitric-oxide synthase. Development of novel 4-oxo-pteridine antagonists of (6R)-5,6,7,8-tetrahydrobiopterin." Kotsonis P., Frohlich L.G., Raman C.S., Li H., Berg M., Gerwig R., Groehn V., Kang Y., Al-Masoudi N., Taghavi-Moghadam S., Mohr D., Munch U., Schnabel J., Martasek P., Masters B.S.S., Strobel H., Poulos T., Matter H., Pfleiderer W., Schmidt H.H.H.W. J. Biol. Chem. 276:49133-49141(2001) [PubMed: 11590164] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.35 ANGSTROMS).

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M99057 mRNA. Translation: AAA30667.1.
M89952 mRNA. Translation: AAA30494.1.
M95674 mRNA. Translation: AAA30669.1.

IPI

IPI00712930.

PIR

A38943.

RefSeq

NP_851380.1.

UniGene

Bt.4662

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1D0C	X-ray	1.65	A/B	39-482	[»]
1D0O	X-ray	1.95	A/B	39-482	[»]
1D1V	X-ray	1.93	A/B	39-482	[»]
1D1W	X-ray	2.00	A/B	39-482	[»]
1D1X	X-ray	2.00	A/B	39-482	[»]
1D1Y	X-ray	2.20	A/B	39-482	[»]
1DM6	X-ray	1.95	A/B	39-482	[»]
1DM7	X-ray	2.10	A/B	39-482	[»]
1DM8	X-ray	2.25	A/B	39-482	[»]
1DMI	X-ray	2.00	A/B	39-482	[»]
1DMJ	X-ray	2.35	A/B	39-482	[»]
1DMK	X-ray	1.90	A/B	39-482	[»]
1ED4	X-ray	1.86	A/B	39-482	[»]
1ED5	X-ray	1.80	A/B	39-482	[»]
1ED6	X-ray	2.05	A/B	39-482	[»]
1FOI	X-ray	1.93	A/B	39-482	[»]
1FOJ	X-ray	2.10	A/B	39-482	[»]
1FOL	X-ray	2.20	A/B	39-482	[»]
1FOO	X-ray	2.00	A/B	39-482	[»]
1FOP	X-ray	2.30	A/B	39-482	[»]
1I83	X-ray	2.00	A/B	39-482	[»]
1NSE	X-ray	1.90	A/B	39-482	[»]
1P6L	X-ray	2.35	A/B	67-482	[»]
1P6M	X-ray	2.27	A/B	67-482	[»]
1P6N	X-ray	2.50	A/B	67-482	[»]
1Q2O	X-ray	1.74	A/B	67-481	[»]
1RS8	X-ray	2.30	A/B	67-481	[»]
1RS9	X-ray	2.22	A/B	67-481	[»]
1ZZS	X-ray	1.85	A/B	67-481	[»]
1ZZT	X-ray	2.14	A/B	67-481	[»]
2G6O	X-ray	1.90	A/B	67-481	[»]
2HX2	X-ray	1.95	A/B	67-481	[»]
2NSE	X-ray	2.34	A/B	39-482	[»]
3DQS	X-ray	2.03	A/B	67-482	[»]
3DQT	X-ray	2.54	A/B	67-482	[»]
3E7S	X-ray	2.50	A/B	57-487	[»]
3NSE	X-ray	2.10	A/B	39-482	[»]
4NSE	X-ray	1.95	A/B	39-482	[»]
5NSE	X-ray	1.90	A/B	39-482	[»]
6NSE	X-ray	2.35	A/B	39-482	[»]
7NSE	X-ray	2.35	A/B	39-482	[»]
8NSE	X-ray	2.25	A/B	39-482	[»]
9NSE	X-ray	2.24	A/B	39-482	[»]

SMR

P29473. Positions 509-1162, 731-1164.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-42217N.

STRING

P29473.

Genome annotation databases

Ensembl

ENSBTAT00000023515; ENSBTAP00000023515; ENSBTAG00000017680; Bos taurus. [Genome view]

GeneID

287024.

KEGG

bta:287024.

Organism-specific databases

CTD

287024.

Phylogenomic databases

eggNOG

maNOG14061.

HOVERGEN

P29473.

InParanoid

P29473.

PhylomeDB

P29473.

Enzyme and pathway databases

BRENDA

1.14.13.39. 251.

Family and domain databases

InterPro

IPR003097. FAD-binding_1.
IPR017927. Fd_Rdtase_FAD-bd.
IPR001094. Flavdoxin-like.
IPR008254. Flavodoxin/NO_synth.
IPR001709. Flavoprot_Pyr_Nucl_cyt_Rdtase.
IPR012144. Nitric-oxide_synthase.
IPR004030. NO_synthase_oxygenase_dom.
IPR001433. OxRdtase_FAD/NAD_bd.
IPR017938. Riboflavin_synthase-like_b-brl.
[Graphical view]

Gene3D

G3DSA:3.90.340.10. NO_synthase_oxygenase_reg. 1 hit.

Pfam

PF00667. FAD_binding_1. 1 hit.
PF00258. Flavodoxin_1. 1 hit.
PF00175. NAD_binding_1. 1 hit.
PF02898. NO_synthase. 1 hit.
[Graphical view]

PIRSF

PIRSF000333. NOS. 1 hit.

PRINTS

PR00369. FLAVODOXIN.
PR00371. FPNCR.

PROSITE

PS51384. FAD_FR. 1 hit.
PS50902. FLAVODOXIN_LIKE. 1 hit.
PS60001. NOS. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

NOS3_BOVIN

Accession

Primary (citable) accession number: P29473

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 1, 1993
Last sequence update:	January 23, 2007
Last modified:	January 19, 2010
This is version 123 of the entry and version 3 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)