TABLE 1

The myometrial nitrosylproteome

A total of 118 unique proteins were identified, 62 of which have not been previously described as being S-nitrosylated, 75 proteins were disparately S-nitrosylated only during pregnancy and 10 were disparately S-nitrosylated only during non-pregnancy, and 33 proteins were S-nitrosylated in both pregnancy and the nonpregnant tissue.

Protein	Biological Process	Previously Identified	NP	P	UniProt	No. of Peptides	Coverage
							%
40S ribosomal protein S11^{^a}	Protein binding	Y C131 (PMID: 20585580)	Y	Y	P62280	3	22.8
40S ribosomal protein S28^{^a}	Protein binding	N	Y	N	P62857	2	33
4-Trimethylamino-butyraldehyde dehydrogenase^{^a}	Metabolism	Y C(74, 173, 443) (PMID: 20585580)	N	Y	P49189	3	8.65
60-kDa Heat shock protein, mitochondrial^{^a}	Protein folding	Y C(237, 442, 447) (PMID: 20585580)	N	Y	P10809	2	9.95
60S ribosomal protein L8^{^a}	rRNA binding	N	Y	N	P62917	2	12.7
6-Phosphogluconate dehydrogenase, decarboxylation^{^a}	Metabolism	N	N	Y	P52209	2	11.88
Actin, α skeletal muscle^{^a}	Cytoskeletal	Y (PMID: 10961840)	Y	Y	P68133	4	13.3
Actin, cytoplasmic 1^{^a}	Cytoskeletal	Y C(217, 272, 285) (PMID: 20585580)	Y	Y	P60709	29	76.13
ADP-ribosylation factor 4^{^a}	Transport	N	N	Y	P18085	3	36.67
Aflatoxin B1 aldehyde reductase member 2^{^a}	Metabolism	N	N	Y	O43488	2	10.46
Alcohol dehydrogenase [NADP⁺]^{^a}	Metabolism	Y (PMID: 19738628)	N	Y	P14550	5	28.62
Aldehyde dehydrogenase, mitochondrial^{^a}	Metabolism	Y C66 (PMID: 20585580)	N	Y	P05091	2	8.83
Aldose reductase^{^a}	Metabolism	Y C299 (PMID: 20585580)	N	Y	P15121	3	28.48
α-2-HS-glycoprotein^{^a}	Endocytosis	N	Y	N	P02765	2	10.61
α-Actinin-1^{^a,^b}	Cytoskeletal	Y C332 (PMID: 20585580)	N	Y	P12814	4	7.85
α-Aminoadipic semialdehyde dehydrogenase^{^a}	Metabolism	N	N	Y	P49419	2	10.57
α-Enolase^{^a}	Metabolism	Y C(119,357) (PMID: 20585580)	Y	Y	P06733	11	34.79
Ankyrin-3^{^a}	Cytoskeletal membrane linker	Y C(1123, 1246, 1369, 2096, 2835, 3395, 3958, 3980, 4233, 4321) (PMID: 20585580)	Y	N	Q12955	2	0.25
Annexin A1^{^a}	Exocytosis and phospholipase A₂ regulation	N	N	Y	P04083	8	35.84
Annexin A2^{^a,^b}	Cytoskeletal adapter	Y (PMID: 12199706)	N	Y	P07355	16	53.69
Annexin A3^{^a,^b}	Inhibitor of phospholipase A₂	N	N	Y	P12429	5	23.53
Annexin A4^{^a}	Exocytosis, membrane fusion	N	N	Y	P09525	2	14.33
Annexin A5^{^a}	Blood coagulation	N	N	Y	P08758	2	11.29
ATP synthase subunit α, mitochondrial^{^a}	Transport	Y (PMID: 20585580)	N	Y	P25705	2	4.73
Calpain small subunit 1^{^a}	Cytoskeletal remodeling and signal transduction	N	N	Y	P04632	4	34.98
Calponin-1^{^a}	Smooth muscle contraction; interaction of calponin with actin inhibits actomyosin Mg-ATPase	N	Y	N	P51911	2	17.67
Cell division cycle 42 GTP binding protein, 25 kDa^{^a}	Cell division	N	N	Y	Q5JYX0	5	28.8
Chloride intracellular channel protein 1^{^a}	Ion transport (redox regulated)	N	N	Y	O00299	2	18.26
Cofilin-1^{^a}	Cytoskeletal actin binding	N	Y	Y	P23528	6	56.02
Corticosteroid-binding globulin^{^a}	Transport	N	N	Y	P08185	2	14.07
Creatine kinase B-type^{^a}	Metabolism	Y (PMID: 18670085)	N	Y	P12277	8	50.66
Cysteine- and glycine-rich protein 1^{^a}	Zinc ion binding	N	Y	Y	P21291	2	27.98
Cysteine sulfinic acid decarboxylase^{^a}	Metabolism	N	N	Y	Q9Y600	5	16.53
Cysteine-rich protein 2^{^a}	Zinc ion binding; interacts with TGFB1I1	N	Y	Y	P52943	7	7.5
Cytochrome c oxidase subunit 2^{^b}	Electron transport	Y (PMID: 15561762)	N	Y	P00403	4	19.38
Desmin^{^a,^b}	Cytoskeletal	Y C333 (PMID: 20585580)	Y	Y	P17661	18	50.21
Destrin^{^a,^b}	Actin depolymerization	Y (PMID: 18670085)	N	Y	P60981	9	46.06
							%
Dihydropyrimidinase-like 2^{^a}	Hydrolase	N	N	Y	Q86U75	2	11.07
Dihydropyrimidinase-related protein 2^{^a}	Cell signaling	Y C504 (PMID: 16418269)	Y	N	Q16555	2	3.03
Elongation factor 1-α1^{^a}	Protein biosynthesis	Y C(3234, 411) (PMID: 20585580)	N	Y	P68104	4	21.54
Elongation factor 1-γ^{^a}	Protein biosynthesis	Y C194 (PMID: 20585580)	N	Y	P26641	2	11.67
Elongation factor 2^{^a}	Translation elongation	Y C(41, 567) (PMID: 20585580)	Y	Y	P13639	7	15.02
Eukaryotic initiation factor 4A-I^{^a}	Protein biosynthesis	N	N	Y	P60842	5	19.6
Fatty acid-binding protein, epidermal^{^a}	Transport	N	N	Y	Q01469	2	15.56
Filamin-A^{^a}	Cytoskeletal scaffold actin binding	Y C717 (PMID: 20585580)	Y	Y	P21333	33	19.74
Filamin-C^{^a}	Cytoskeletal actin cross-linker	Y C (805, 1680, 2096, 2660) (PMID: 20585580)	N	Y	Q14315	10	6.93
Galectin-1^{^a}	Regulation of apoptosis, cell proliferation/differentiation	Y C61 (PMID: 20585580)	N	Y	P09382	5	39.26
Glutaredoxin-1^{^a}	Transport and thiol reduction	Y (PMID: 17355958)	N	Y	P35754	2	38.68
Glutathione transferase Mu 2^{^a}	Metabolism	Y (PMID: 20585580)	N	Y	P28161	5	23.96
Glutathione transferase Mu 3^{^a}	Metabolism	Y C3 (PMID: 20585580)	N	Y	P21266	2	16.59
Glutathione transferase P^{^a}	Metabolism	Y C(47, 101) (PMID: 11533048)	Y	Y	P09211	8	59.52
Glyceraldehyde-3-phosphate dehydrogenase^{^a,^b}	Metabolism	Y C(152, 156, 247) (PMID: 20585580)	N	Y	P04406	12	59.46
Glycerol-3-phosphate dehydrogenase 1-like protein^{^a}	Regulation of sodium current	N	N	Y	Q8N335	2	10.8
G_i subunit α-2^{^a}	Signal transduction	N	N	Y	P04899	2	9.58
Heat shock 105 kDa/110 kDa protein 1^{^a}	Chaperone	N	N	Y	Q5TBM3	2	4.42
Heat shock protein 90 kDa α cytosolic, class B member 1^{^a}	Chaperone	N	Y	N	Q5T9W8	2	5.75
Heat shock protein β-1^{^a,^b}	Cytoskeletal remodeling and signal transduction	Y (PMID: 18670085)	Y	Y	P04792	11	84.5
Heat shock protein β-6^{^a}	Stress response (HSP20 family)	N	N	Y	O14558	4	51.85
Heat shock protein HSP90-α^{^b}	Chaperone	Y C(481, 597, 598) (PMID: 20585580)	Y	Y	P07900	10	14
Hemoglobin subunit α^{^a,^b}	Oxygen transport	Y C105 (PMID: 20585580)	Y	Y	P69905	8	57.04
Hemoglobin subunit β^{^a,^b}	Oxygen transport	Y C93 (PMID: 16594178)	Y	Y	P68871	13	90.48
Histidine-rich glycoprotein^{^a}	Blood coagulation	N	N	Y	P04196	2	3.68
Kinesin family member 17^{^a}	Microtubule-based movement	N	N	Y	A2A3Q7	2	4.91
Lipoma-preferred partner^{^a}	Cell signaling and protein scaffold	N	Y	Y	Q93052	14	31.97
l-Lactate dehydrogenase A chain^{^a}	Metabolism	Y C163 (PMID: 20585580)	N	Y	P00338	10	54.14
Malate dehydrogenase, cytoplasmic^{^b}	Metabolism	Y C(137, 154) (PMID: 20585580)	Y	Y	P40925	12	194 Mowse^{^c}
Myosin light chain kinase, smooth muscle^{^a}	Smooth muscle contraction	N	N	Y	Q15746	3	3.45
Myosin light polypeptide 6^{^a,^b}	Smooth muscle contraction	? (PMID: 20585450)	Y	Y	P60660	9	51.66
Myosin regulatory light polypeptide 9^{^a}	Smooth muscle contraction	? (PMID: 20585450)	N	Y	P24844	4	45.35
Myosin-11^{^a}	Smooth muscle contraction	? (PMID: 20585450)	N	Y	P35749	6	5.27
Neutral α-glucosidase AB^{^b}	Metabolism	N	Y	Y	Q14697	4	3.91
Nucleoside diphosphate kinase A^{^a}	Metabolism and cell signaling	N	N	Y	P15531	2	25.47
Oligoribonuclease, mitochondrial^{^a}	Exonuclease	N	N	Y	Q9Y3B8	2	18.2
PDZ and LIM domain protein 1^{^a}	Cytoskeletal adapter (interacts with α-actinins 1, 2, and 4)	N	N	Y	O00151	3	19.82
Peptidyl-prolyl cis-trans isomerase A^{^a}	Protein folding	N	Y	Y	P62937	7	51.52
Peroxiredoxin-1^{^a}	Redox regulation	Y C(52, 173) (PMID: 20585580)	Y	Y	Q06830	6	36.14
Phosphofructokinase, platelet^{^a}	Metabolism	N	N	Y	Q5VSR7	3	4.8
Phosphoglucomutase-like protein 5^{^a}	Cytoskeletal; interacts with cytoskeletal proteins dystrophin and utrophin	N	N	Y	Q15124	2	7.14
Phosphoglycerate kinase 1^{^a}	Metabolism	Y C50 (PMID: 20585580)	N	Y	P00558	9	42.82
Phosphoglycerate mutase 1^{^a}	Metabolism	Y C55 (PMID: 20585580)	N	Y	P18669	2	13.39
Prefoldin subunit 3^{^a}	Protein folding	N	N	Y	P61758	2	28.43
Preproalbumin precursor^{^a}	Guinea pig specific	N	Y	Y	Q6WDN9	39	65.46
							%
Profilin-1^{^a}	Cytoskeletal remodeling	Y (PMID: 18283105)	N	Y	P07737	3	22.86
Prolow-density lipoprotein receptor-related protein 1^{^a}	Endocytosis and cell signaling	N	N	Y	Q07954	2	0.9
Prorelaxin H1^{^a}	Signal transduction and tissue remodeling	N	N	Y	P04808	3	51.25
Protein DJ-1^{^a}	Transcriptional activator	N	N	Y	Q99497	5	38.62
Protein S100-A10^{^a}	Protein binding; induces the dimerization of ANXA2/p36	N	N	Y	P60903	2	37.5
Protein S100-A11^{^a,^b}	Calcium binding and signal transduction	Y C13 (PMID: 20585580)	N	Y	P31949	7	53.92
Protein S100-A4^{^a,^b}	Calcium binding	N	N	Y	P26447	2	11.88
Protein S100-A9^{^a}	Signal transduction	Y (PMID: 18832721)	N	Y	P06702	3	51.26
Purine nucleoside phosphorylase^{^b}	NAD biosynthesis	N	N	Y	P00491	4	28.82
Pyruvate kinase isozymes M1/M2^{^a}	Metabolism	Y C(326, 358, 423, 474) (PMID: 20585580)	Y	Y	P14618	14	47.27
Ras-related protein Rab-1A^{^a}	Transport	N	N	Y	P62820	3	35.12
Ras-related protein R-Ras^{^a}	Cytoskeletal remodeling	N	N	Y	P10301	3	26.15
Serotransferrin^{^a}	Transport	Y C(58, 67, 137, 246, 387, 396, 469, 596, 615) (PMID: 20585580)	Y	Y	P02787	6	10.08
Serpin peptidase inhibitor, clade B ovalbumin, memb0.6^{^a}	Peptidase inhibitor	N	N	Y	Q5TD02	11	48.99
S-Formylglutathione hydrolase^{^a}	Metabolism	N	N	Y	P10768	2	15.6
SH3 domain-binding glutamic acid-rich-like protein^{^b}	SH3 binding	No Cys in Human	Y	Y	O75368	2	17.2
Stress-70 protein, mitochondrial^{^a}	Chaperone	Y C487 (PMID: 20585580)	N	Y	P38646	2	4.28
Talin-1^{^a}	Cytoskeletal membrane connector	N	Y	Y	Q9Y490	13	10.27
Testis derived transcript 3 LIM domains^{^a}	Zinc ion binding	N	N	Y	A4D0U5	6	18.89
Thioredoxin^{^b}	S-Nitrosylation regulation	Y (62, 69, 73) (PMID: 20585580)	Y	Y	P10599	7	32
Transforming growth factor β-1-induced transcript 1 protein^{^a}	Molecular adapter at focal adhesion complexes	N	Y	Y	O43294	3	13.3
Transforming protein RhoA^{^a}	Signal transduction	N	N	Y	P61586	2	13.99
Transgelin^{^a,^b}	Actin binding	N	N	Y	Q01995	10	59.7
Transgelin-2^{^a}	Protein binding	N	Y	Y	P37802	7	40.09
Transitional endoplasmic reticulum ATPase^{^a,^b}	Transport	Y C(77, 209, 695) (PMID: 20585580)	Y	N	P55072	2	11
Transthyretin^{^a}	Transport	Y (PMID: 16101296)	Y	N	P02766	2	26.19
Tropomyosin α-4 chain^{^a}	Smooth muscle contraction is regulated by interaction with caldesmon	Y C170 (PMID: 18992711)	N	Y	P67936	3	16.53
Tropomyosin β chain^{^a,^b}	Smooth muscle contraction is regulated by interaction with caldesmon	Y C170 (PMID: 19447776)	Y	Y	P07951	4	29.58
Tryptophanyl-tRNA synthetase, cytoplasmic^{^a}	Protein biosynthesis	N	N	Y	P23381	2	12.13
Tubulin β chain^{^a,^b}	Cytoskeletal	Y (PMID: 16418269)	Y	Y	P07437	18	54.05
Tubulin β-2B chain^{^a}	Cytoskeletal	N	N	Y	Q9BVA1	15	46.07
Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, β polypeptide^{^a}	Protein binding	N	N	Y	Q59EQ2	2	24.39
Vimentin^{^a,^b}	Cytoskeletal remodeling	Y C328 (PMID: 20585580)	Y	Y	P08670	18	50.22
Vinculin^{^a}	Cytoskeletal actin binding	Y (PMID: 17704302)	N	Y	P18206	3	5.11
Vitamin D-binding protein^{^a}	Transport	N	Y	N	P02774	2	5.88
Zinc finger protein 641^{^a}	Transcriptional activator	N	Y	Y	Q96N77	13	40.69

NP, nonpregnant estrogen-primed guinea pig; P, pregnant near term 60- to 67-day timed pregnant; Y, yes; N, no; PMID, PubMed identification number.
↵a Proteins were identified by LTQ-Orbitrap LC-MS/MS.
↵b Proteins were identified by MALDI TOF/TOF.
↵c As identified by spot cutting of Nitrosyl-DIGE gels and MALDI/TOF/TOF analysis.