Literature DB >> 31417950

Cellular proteome datasets of human endothelial cells under physiologic state and after treatment with caffeine and epigallocatechin-3-gallate.

Chanettee Chanthick1, Visith Thongboonkerd1.   

Abstract

Human endothelial cells play several significant roles in vascular biology and homeostasis. We report herein cellular proteome datasets of EA.hy926 human endothelial cells under physiologic condition and after treatment with 100 μM caffeine or EGCG for 24-h. Cellular proteins were extracted and subjected to in-solution tryptic digestion using filter-aided sample preparation (FASP) method. The digested peptides were analyzed by nanoflow liquid chromatography coupled to tandem mass spectrometry (nanoLC-ESI-Qq-TOF MS/MS). Finally, the mass spectral data were searched against the human Swiss-Prot database using Mascot 2.4 search engine and quantified using Skyline v.3.5 software and BiblioSpec algorithm. All of these data were used for further comparative proteomics study followed by bioinformatics analyses to investigate differential biochemical effects of caffeine and EGCG on human endothelial cells (Chanthick et al., 2019) [1].

Entities:  

Keywords:  Cellular response; Coffee; Epigallocatechin-3-gallate; Polyphenol; Proteome; Tea

Year:  2019        PMID: 31417950      PMCID: PMC6690735          DOI: 10.1016/j.dib.2019.104292

Source DB:  PubMed          Journal:  Data Brief        ISSN: 2352-3409


Specifications Table These data are the valuable resource for understanding and further analyzing the cellular proteome of human endothelial cells under physiologic condition and after treatment with caffeine or EGCG. Proteomists, biochemists, physiologists, cellular biologists, and nutritionists will be benefited from these data. Further functional investigations on these data will lead to better understanding and provide significant insights into vascular cell biology.

Data

We report herein the complete lists of proteins identified from EA.hy926 human endothelial cells under physiologic (control) condition (Table 1), after 24-h treatment with caffeine (Table 2), and after 24-h treatment with EGCG (Table 3). Their identities, gene symbols, MS/MS identification scores, percentages of sequence coverage, numbers of distinct and total matched peptides, isoelectric points (pI), and intensity data are shown. All of these data were used for further comparative proteomics study followed by bioinformatics analyses to investigate differential biochemical effects of caffeine and EGCG on human endothelial cells [1].
Table 1

All proteins identified in control EA.hy926 endothelial cells.

Protein nameSwiss-Prot IDGene symbolMS/MS identification score% CovNo. of distinct/total matched peptidespIIntensity (x103 arbitrary unit)
N1N2N3MeanSEM
10 kDa heat shock protein, mitochondrialP61604HSPE14429.43/58.917446395317914131044578
14-3-3 protein beta/alphaP31946YWHAB14720.74/74.8627424019933195453798914
14-3-3 protein epsilonP62258YWHAE13616.53/74.6107271619014735138841634
14-3-3 protein thetaP27348YWHAQ809.42/24.7290784093131645338853600
14-3-3 protein zeta/deltaP63104YWHAZ11441.28/194.716087171161765016951459
26S protease regulatory subunit 6AP17980PSMC3707.72/65.1341934490932290371313928
26S protease regulatory subunit 7P35998PSMC2753.51/25.7108812421519907183343929
26S proteasome non-ATPase regulatory subunit 12O00232PSMD12318.33/47.5191984608929657316487826
3-ketoacyl-CoA thiolase, mitochondrialP42765ACAA2416.51/28.3NANANANANA
40S ribosomal protein S10P46783RPS103614.52/210.2NANANANANA
40S ribosomal protein S11P62280RPS116419.63/510.33489264921482762391
40S ribosomal protein S12P25398RPS12123283/76.85989922323754129895463
40S ribosomal protein S13P62277RPS1314718.53/710.54685651450825427555
40S ribosomal protein S14P62263RPS141237.31/210.1121721889313314147932076
40S ribosomal protein S15P62841RPS1513036.63/510.433753785776349741399
40S ribosomal protein S16P62249RPS1620821.93/610.2183283778615543238866996
40S ribosomal protein S18P62269RPS18448.62/411.0367335093717363363
40S ribosomal protein S19P39019RPS196121.43/510.3318714858640790404164829
40S ribosomal protein S2P15880RPS29917.13/610.3234142533332355270342718
40S ribosomal protein S20P60866RPS208110.11/410.0154791156610693125791471
40S ribosomal protein S25P62851RPS255719.22/310.13224203913502204547
40S ribosomal protein S28P62857RPS284934.82/210.738164070718050221082
40S ribosomal protein S3P23396RPS35710.72/49.7541510341669474831476
40S ribosomal protein S4, X isoformP62701RPS4X3715.63/310.244238517582062531202
40S ribosomal protein S5P46782RPS576317.64/219.7817839583967353816588223
40S ribosomal protein S6P62753RPS65813.33/610.93259266967849126017171
40S ribosomal protein S7P62081RPS713919.63/710.138988383537958871319
40S ribosomal protein S8P62241RPS86110.62/410.34996767760096227782
40S ribosomal protein SAP08865RPSA9818.63/64.8633413254595485142372
60 kDa heat shock protein, mitochondrialP10809HSPD1783239/275.73697386169866296992316476
60S acidic ribosomal protein P0-likeQ8NHW5RPLP0P67814.23/65.4NANANANANA
60S acidic ribosomal protein P1P05386RPLP138141/24.3269015160238425389767136
60S acidic ribosomal protein P2P05387RPLP27510.41/44.4179801062214169142572124
60S ribosomal protein L10P27635RPL1013416.43/410.1NANANANANA
60S ribosomal protein L10aP62906RPL10A4520.74/59.925064238882281223922650
60S ribosomal protein L12P30050RPL1212137.64/69.57612640317450104893498
60S ribosomal protein L13P26373RPL13304.71/211.79680154376803106402538
60S ribosomal protein L18Q07020RPL1810614.42/511.787484197687766071321
60S ribosomal protein L19P84098RPL1910213.32/411.58786945599898017442314423
60S ribosomal protein L23P62829RPL23118252/310.5200561051913973148502788
60S ribosomal protein L23aP62750RPL23A4913.52/410.45572128971082197632180
60S ribosomal protein L27P61353RPL27316.61/210.616193189517547118785007
60S ribosomal protein L30P62888RPL308413.91/29.7588937749466693010312377
60S ribosomal protein L31P62899RPL312518.41/110.5NANANANANA
60S ribosomal protein L38P63173RPL389535.72/810.1NANANANANA
60S ribosomal protein L6Q02878RPL6308.73/410.64941457366165377629
6-phosphogluconate dehydrogenase, decarboxylatingP52209PGD272.31/106.8NANANANANA
78 kDa glucose-regulated proteinP11021HSPA590329.418/345.1NANANANANA
Acidic leucine-rich nuclear phosphoprotein 32 family member AP39687ANP32A718.41/44.0NANANANANA
Actin, aortic smooth muscleP62736ACTA218523111/2065.2NANANANANA
Actin, cytoplasmic 1P60709ACTB1760750.119/13135.3129561515878211591547149166198029
Activator of 90 kDa heat shock protein ATPase homolog 1O95433AHSA1929.52/35.4NANANANANA
AdenosylhomocysteinaseP23526AHCY359.73/45.9179801062214169142572124
ADP/ATP translocase 2P05141SLC25A55110.13/79.7333035281147782446325848
ADP-ribosylation factor 1P84077ARF13617.73/36.3NANANANANA
Alpha-actinin-1P12814ACTN11279.56/85.3NANANANANA
Alpha-actinin-4O43707ACTN430611.37/105.33334498134980637651421585
Alpha-enolaseP06733ENO1168348.414/2977.016184928470022461622372235467
Annexin A1P04083ANXA19373910/316.611524519185415510015406622121
Annexin A2P07355ANXA2199557.818/647.6NANANANANA
Annexin A4P09525ANXA44010.33/35.8329214483230964362394334
Annexin A5P08758ANXA5126938.812/414.921101735982931622029568944168
ATP synthase subunit alpha, mitochondrialP25705ATP5F1A3957.43/99.24459213260513845310521730359
ATP synthase subunit beta, mitochondrialP06576ATP5F1B4462710/545.3297783848027963320743246
ATP-dependent RNA helicase AQ08211DHX9523.64/96.469674726352850741008
ATP-dependent RNA helicase DDX1Q92499DDX1692.81/26.8NANANANANA
Barrier-to-autointegration factorO75531BANF111429.22/35.8NANANANANA
Basic leucine zipper and W2 domain-containing protein 1Q7L1Q6BZW11639.83/45.8NANANANANA
Beta-enolaseP13929ENO3109026.56/1347.6NANANANANA
CalnexinP27824CANX1442.51/34.5NANANANANA
Calpain small subunit 1P04632CAPNS18591/25.1NANANANANA
CalreticulinP27797CALR54123/44.35250507764655598437
Casein kinase II subunit alpha 3Q8NEV1CSNK2A336102/48.5NANANANANA
Cathepsin DP07339CTSD1248.52/36.12319321425992711264
CD9 antigenP21926CD979111/26.82925331242693502399
Cell division control protein 42 homologP60953CDC4223814.12/126.23293874878436605049212580
Chloride intracellular channel protein 1O00299CLIC11137.51/25.1214642830621295236892309
Clathrin heavy chain 1Q00610CLTC1346.17/125.5NANANANANA
Cleavage and polyadenylation specificity factor subunit 6Q16630CPSF6645.82/36.7NANANANANA
Coatomer subunit deltaP48444ARCN1356.32/25.9742265113981597560
Cofilin-1P23528CFL11046.61/28.2123921582920905163752472
Cofilin-2Q9Y281CFL26011.42/37.7NANANANANA
Copine-1Q99829CPNE18910.24/115.58553885269498118591
Cysteine and glycine-rich protein 1P21291CSRP1448.81/58.9NANANANANA
Cysteine and histidine-rich domain-containing protein 1Q9UHD1CHORDC1304.51/18.1NANANANANA
Cytoplasmic dynein 1 heavy chain 1Q14204DYNC1H1432.510/116.0NANANANANA
Cytoskeleton-associated protein 4Q07065CKAP434521.910/185.6304153440831530321181189
Dedicator of cytokinesis protein 10Q96BY6DOCK10392.77/276.7300143249938295336032453
Dihydropyrimidinase-related protein 2Q16555DPYSL2305.82/26.0NANANANANA
DNA damage-binding protein 1Q16531DDB11264.63/45.1NANANANANA
DNA-(apurinic or apyrimidinic site) lyaseP27695APEX1835.31/28.35111397763275138678
Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1P04843RPN1327.64/336.089665549370860741540
E3 ubiquitin/ISG15 ligase TRIM25Q14258TRIM25362.91/18.4NANANANANA
EH domain-containing protein 1Q9H4M9EHD13110.53/56.43632359544283885272
EH domain-containing protein 2Q9NZN4EHD2907.23/46.02614193224952347210
Elongation factor 1-alpha 1P68104EEF1A1209024.56/1969.11878353104480053996410692
Elongation factor 1-deltaP29692EEF1D9012.82/24.95966786763346722582
Elongation factor 2P13639EEF290727.515/356.411594925441223204720080242915
Elongation factor Tu, mitochondrialP49411TUFM12915.34/67.37993715410111221111541621469
EndoplasminP14625HSP90B132516.211/214.8NANANANANA
Enoyl-CoA hydratase, mitochondrialP30084ECHS118413.83/88.3109778799606986151420
Eukaryotic initiation factor 4A-IP60842EIF4A114414.54/65.3748712602897896891519
Eukaryotic translation initiation factor 3 subunit BP55884EIF3B321.71/14.93901330516912965660
Eukaryotic translation initiation factor 3 subunit EP60228EIF3E316.53/45.7NANANANANA
Eukaryotic translation initiation factor 3 subunit HO15372EIF3H645.41/26.16881606565226491525
Eukaryotic translation initiation factor 4BP23588EIF4B402.81/15.6NANANANANA
EzrinP15311EZR25311.16/135.96525723211140783418215759
FACT complex subunit SSRP1Q08945SSRP1747.64/66.5NANANANANA
F-actin-capping protein subunit alpha-1P52907CAPZA1436.31/25.5NANANANANA
FascinQ16658FSCN1244.91/16.8NANANANANA
Filamin-BO75369FLNB1754.99/135.5439593964849052442202718
Fructose-bisphosphate aldolase AP04075ALDOA60236.810/258.3139031540934490212676626
Galectin-1P09382LGALS131628.94/95.3321435794145901453287453
Glucose-6-phosphate isomeraseP06744GPI9610.23/78.48434916231414132012932823292
Glucosidase 2 subunit betaP14314PRKCSH677.84/84.36655620392887382962
Glutathione S-transferase PP09211GSTP139941.94/115.410280130531259611976858
Glyceraldehyde-3-phosphate dehydrogenaseP04406GAPDH630654.615/5118.639311361797048702449936965203
Glycerol kinase 2Q14410GK2524.52/45.6NANANANANA
GTP-binding nuclear protein RanP62826RAN6910.62/47.014089134871342913668211
HEAT repeat-containing protein 5AQ86XA9HEATR5A320.41/2096.110876614341910289811836112643
Heat shock 70 kDa protein 6P17066HSPA62818.25/85.8491917851360021625758560
Heat shock cognate 71 kDa proteinP11142HSPA8103227.618/415.4NANANANANA
Heat shock protein beta-1P04792HSPB122932.75/266.052530104183974078470716207
Heat shock protein HSP 90-alphaP07900HSP90AA179024.917/384.919339938638530257229411955870
Heat shock protein HSP 90-betaP08238HSP90AB166425.816/385.018762330258528571125864035841
Heterogeneous nuclear ribonucleoprotein A1P09651HNRNPA1260184/139.2128652121015315164632476
Heterogeneous nuclear ribonucleoprotein A3P51991HNRNPA3498.72/39.1NANANANANA
Heterogeneous nuclear ribonucleoprotein D0Q14103HNRNPD8816.96/107.6527711439541273762032
Heterogeneous nuclear ribonucleoprotein HP31943HNRNPH128112.54/105.927089295662899328550749
Heterogeneous nuclear ribonucleoprotein H3P31942HNRNPH3884.91/26.422117723302643201717
Heterogeneous nuclear ribonucleoprotein KP61978HNRNPK7011.24/55.457721416310690102082434
Heterogeneous nuclear ribonucleoprotein LP14866HNRNPL333.11/18.5NANANANANA
Heterogeneous nuclear ribonucleoprotein MP52272HNRNPM477.14/78.88039245089211139195305
Heterogeneous nuclear ribonucleoprotein QO60506SYNCRIP825.33/78.71793966972511844536514450
Heterogeneous nuclear ribonucleoprotein RO43390HNRNPR1434.12/78.278839067104659138746
Heterogeneous nuclear ribonucleoprotein UQ00839HNRNPU1274.73/65.8297262760385352367410762
Heterogeneous nuclear ribonucleoproteins A2/B1P22626HNRNPA2B1248113/99.0115551548215807142821366
Heterogeneous nuclear ribonucleoproteins C1/C2P07910HNRNPC785.92/55.0NANANANANA
High mobility group protein B1P09429HMGB1454.21/25.68243772086168193260
High mobility group protein HMG-I/HMG-YP17096HMGA15122.42/310.3866661501213989851736
High mobility group protein HMGI-CP52926HMGA284333/710.6103731346015939132571610
Histone H1.0P07305H1F0395.21/110.83673477436074018379
Histone H1.4P10412HIST1H1E61162/311.0711810081497573911480
Histone H2A type 1-B/EP04908HIST1H2AB77028.53/1711.119358227078121105222513823372
Histone H2A type 1-DP20671HIST1H2AD100028.53/2110.9NANANANANA
Histone H2A type 2-CQ16777HIST2H2AC85327.13/1910.9NANANANANA
Histone H2B type 1-BP33778HIST1H2BB262534.94/10210.3NANANANANA
Histone H2B type 1-C/E/F/G/IP62807HIST1H2BC208135.75/8310.3NANANANANA
Histone H2B type 1-MQ99879HIST1H2BM266440.56/13910.3NANANANANA
Histone H3.1tQ16695HIST3H369323.55/6511.1NANANANANA
Histone H4P62805HIST1H4A125051.57/5411.417307524762426679422916428585
Importin-7O95373IPO7283.13/104.7333982425537762986614929
Importin-9Q96P70IPO9353.22/34.7NANANANANA
Inosine-5′-monophosphate dehydrogenase 2P12268IMPDH2497.44/46.45029671577466497792
Integrin beta-4P16144ITGB43436/95.7798012878908399801483
Kelch-like protein 35Q6PF15KLHL351391.21/808.16734616872311462811689929287
Keratin, type I cytoskeletal 18P05783KRT1843444.215/305.39775116805411808912796520887
Keratin, type II cytoskeletal 1P04264KRT1644.53/38.274625503196549771608
Keratin, type II cytoskeletal 7P08729KRT751124.713/275.410180514336812090712202712011
Keratin, type II cytoskeletal 8P05787KRT86214124/465.517907222829622787521174816338
Lamin-B1P20700LMNB16112.36/65.1837711486858794831003
Leucine--tRNA ligase, cytoplasmicQ9P2J5LARS372.21/27.0NANANANANA
l-lactate dehydrogenase A chainP00338LDHA46634.611/228.48403114031211294411242916249
l-lactate dehydrogenase B chainP07195LDHB78126.38/255.79957816232114509513566518716
Long-chain-fatty-acid--CoA ligase 4O60488ACSL4324.12/38.71776344120312416518
Lysine-specific demethylase 2BQ8NHM5KDM2B335.86/78.9NANANANANA
Macrophage migration inhibitory factorP14174MIF377.81/17.720861890228020163367596
Malate dehydrogenase, mitochondrialP40926MDH21839.52/38.9172373619011453363
Malignant T-cell-amplified sequence 1Q9ULC4MCTS1669.41/29.0112652434319140182493801
Matrin-3P43243MATR3542.82/45.9621983046679695241123503
MoesinP26038MSN70535.719/456.1115884100784689809521613823
Myosin light polypeptide 6P60660MYL614229.13/74.6NANANANANA
Myosin-9P35579MYH967913.821/485.51278078147214950511959520064
Nascent polypeptide-associated complex subunit alpha, muscle-specific formE9PAV3NACA1683.66/89.6800611432602684881579
Neuroblast differentiation-associated protein AHNAKQ09666AHNAK834.923/555.8170423233330396265904807
Neutral alpha-glucosidase ABQ14697GANAB441.41/15.739898636910772441633
Non-POU domain-containing octamer-binding proteinQ15233NONO1054.91/49.0NANANANANA
Nuclear autoantigenic sperm proteinP49321NASP273.62/24.3NANANANANA
NucleolinP19338NCL44618.710/264.64183543031504254659
NucleophosminP06748NPM171925.56/224.65414292885746317388611190
Nucleoside diphosphate kinase AP15531NME113511.21/45.8NANANANANA
Nucleoside diphosphate kinase BP22392NME213923.72/58.5NANANANANA
Nucleosome assembly protein 1-like 1P55209NAP1L120616.15/84.4157982728411672182514671
Obg-like ATPase 1Q9NTK5OLA16210.43/57.68471614116052102212992
ParathymosinP20962PTMS8720.62/44.1NANANANANA
Peptidyl-prolyl cis-trans isomerase AP62937PPIA72062.410/407.724580336299837458132779441132
Peptidyl-prolyl cis-trans isomerase BP23284PPIB26022.74/79.4303523798916548282966274
Peroxiredoxin-1Q06830PRDX121929.15/108.36791723762247263880114560
Peroxiredoxin-6P30041PRDX64112.52/36.02287348237333167446
Phosphoglycerate kinase 1P00558PGK144426.67/188.3268133613026255297323203
Phosphoglycerate mutase 1P18669PGAM1160223/86.7NANANANANA
Plasminogen activator inhibitor 1 RNA-binding proteinQ8NC51SERBP1515.92/38.75170832267306741910
Plasminogen activator inhibitor 2P05120SERPINB2264.61/15.53059513641664121600
Plastin-3P13797PLS3959.74/85.4249693016125580269031638
PlectinQ15149PLEC66212.349/855.724480630766348015734420970354
Poly(rC)-binding protein 2Q15366PCBP2697.42/36.3NANANANANA
Polymerase I and transcript release factorQ6NZI2CAVIN122593/75.5119674997577682323713424
Polypyrimidine tract-binding protein 1P26599PTBP1837.32/69.2NANANANANA
PRA1 family protein 3O75915ARL6IP59619.13/69.8NANANANANA
Prelamin-A/CP02545LMNA93928.218/386.611164220251412802914739527963
Pre-mRNA-splicing factor CWC25 homologQ9NXE8CWC25312.41/210.2317982573241296329424529
Probable ATP-dependent RNA helicase DDX17Q92841DDX17756.95/98.523702019598634581268
Probable ATP-dependent RNA helicase DDX5P17844DDX52166.43/69.1860985143740345592923876
Profilin-1P07737PFN118359.37/118.4238052791714580221013943
ProhibitinP35232PHB353.71/15.64960340929803783601
Prohibitin-2Q99623PHB25515.14/59.83673263234613255318
Proliferating cell nuclear antigenP12004PCNA5420.33/84.61493217224519642804011980
Proteasome activator complex subunit 2Q9UL46PSME26316.73/45.557261591511014108852942
Protein disulfide-isomerase A3P30101PDIA314015.85/86.010505141348601110801623
Protein disulfide-isomerase A4P13667PDIA4486.83/55.024845511753214989390277328
Protein disulfide-isomerase A6Q15084PDIA616016.44/85.0181103261918642231244750
Protein S100-A11P31949S100A11106743.84/296.63188358733834435802014888
Protein S100-A6P06703S100A67116.72/35.3273472483229523272341355
Protein transport protein Sec61 subunit betaP60468SEC61B3541.73/311.6NANANANANA
Protein-glutamine gamma-glutamyltransferase 2P21980TGM244216.48/155.12635543480649304492211159
Prothymosin alphaP06454PTMA15112.62/33.743185735976681940411891
Proto-oncogene serine/threonine-protein kinase mosP00540MOS356.42/39.22580173938653245502114820
Purine nucleoside phosphorylaseP00491PNP2512.83/46.52740575736844060891
Putative nascent polypeptide-associated complex subunit alpha-like proteinQ9BZK3NACAP13171/14.568023279361945671122
Putative Ras-related protein Rab-1CQ92928RAB1C11120.92/95.33926305562674416959
Putative UPF0633 protein MGC21881A6NN06MGC21881327.41/811.9249934196921538295006314
Pyruvate dehydrogenase E1 component subunit beta, mitochondrialP11177PDHB384.51/16.2423653498467573483015420
Pyruvate kinase PKMP14618PKM26256126/2838.043854566697054505055018865991
Rab GDP dissociation inhibitor betaP50395GDI2936.72/56.17791269128837145146207
Ras GTPase-activating-like protein IQGAP1P46940IQGAP1625.77/126.113994811738945331735
Ras-related protein Rab-1BQ9H0U4RAB1B1158.51/45.6NANANANANA
Ras-related protein Rab-5CP51148RAB5C43192/28.66423597634325277932
Receptor of activated protein C kinase 1P63244RACK1417.62/47.62447443245493810682
Rho GDP-dissociation inhibitor 1P52565ARHGDIA1257.41/25.03282523645564358573
Rho GDP-dissociation inhibitor 2P52566ARHGDIB4612.91/15.1NANANANANA
Rho GTPase-activating protein 26Q9UNA1ARHGAP26402.92/86.2NANANANANA
Ribonuclease inhibitorP13489RNH19318.25/74.73381078512509345441913021
RNA polymerase II-associated protein 1Q9BWH6RPAP1292.33/46.0535911227365667472293
RNA-binding motif protein, X chromosomeP38159RBMX4113.84/510.194195591372862461675
RNA-binding protein FUSP35637FUS2627.82/49.4NANANANANA
Serpin H1P50454SERPINH11268.92/68.8210585428140568386359639
SH3 domain-binding glutamic acid-rich-like protein 3Q9H299SH3BGRL339282/34.8NANANANANA
Signal recognition particle 14 kDa proteinP37108SRP1417110.31/210.1NANANANANA
Small nuclear ribonucleoprotein Sm D1P62314SNRPD13416.81/411.6409940358189532113610524
Sorting nexin-6Q9UNH7SNX6355.21/15.8NANANANANA
Splicing factor 3B subunit 1O75533SF3B1314.14/96.7NANANANANA
Splicing factor, proline- and glutamine-richP23246SFPQ1457.23/69.52648224428992597191
StathminP16949STMN13661/15.85573373463205209768
Stress-70 protein, mitochondrialP38646HSPA910814.98/125.9390275602366009536867876
Stress-induced-phosphoprotein 1P31948STIP1345.53/36.422445934200633951272
Sulfotransferase 1A3P0DMM9SULT1A36911.22/45.7NANANANANA
Surfeit locus protein 4O15260SURF4876.71/47.6NANANANANA
T-complex protein 1 subunit betaP78371CCT23318.95/56.0NANANANANA
T-complex protein 1 subunit deltaP50991CCT411010.65/108.04676544424794200888
T-complex protein 1 subunit etaQ99832CCT720715.34/117.6NANANANANA
T-complex protein 1 subunit gammaP49368CCT348154/56.1NANANANANA
T-complex protein 1 subunit thetaP50990CCT81088.83/65.445589106359689747364117697
T-complex protein 1 subunit zetaP40227CCT6A13214.55/86.2288024173731766341023912
Thioredoxin domain-containing protein 5Q8NBS9TXNDC5210134/85.634293877862131614011348
Thioredoxin reductase 1, cytoplasmicQ16881TXNRD1994.82/37.2NANANANANA
TransaldolaseP37837TALDO13152/36.416133864910676118192235
Transcription elongation factor A protein-like 3Q969E4TCEAL337202/64.9NANANANANA
Transcription factor BTF3P20290BTF32510.71/19.4NANANANANA
Transcription intermediary factor 1-betaQ13263TRIM28545.43/55.51251891191238744811058711701
Transgelin-2P37802TAGLN275249.78/268.492336122765748519665113999
Transitional endoplasmic reticulum ATPaseP55072VCP2217.14/205.1339714345235614376792925
TransketolaseP29401TKT60415.48/187.6335905187652238459016157
Triosephosphate isomeraseP60174TPI17815710/265.71766071914032001031893716858
Tropomodulin-3Q9NYL9TMOD3399.72/25.13993661894238511640
Tropomyosin alpha-4 chainP67936TPM414028.67/134.7NANANANANA
Tubulin alpha-1B chainP68363TUBA1B75031.99/314.915433728634722956622341738232
Tubulin beta chainP07437TUBB91625.79/334.810357319197313837014463825711
Tubulin beta-4A chainP04350TUBB4A535238/224.81375217626513450911609819935
Tubulin beta-4B chainP68371TUBB4B60125.210/244.8NANANANANA
Tubulin beta-6 chainQ9BUF5TUBB634823.38/174.8357954377155976451815868
Ubiquitin carboxyl-terminal hydrolase isozyme L1P09936UCHL16236.85/55.3117263051824641222955550
Ubiquitin-40S ribosomal protein S27aP62979RPS27A34031.44/129.7NANANANANA
Ubiquitin-like modifier-activating enzyme 1P22314UBA12805.94/95.5252913204434487306072750
UMP-CMP kinaseP30085CMPK13417.92/35.4NANANANANA
UPF0183 protein C16orf70Q9BSU1C16orf70326.62/57.6330645434436724413776569
UPF0258 protein KIAA1024Q9UPX6KIAA10243643/67.0799112747235976993003
Vesicle-trafficking protein SEC22bO75396SEC22B4610.22/36.4NANANANANA
VimentinP08670VIM315959.430/2965.16255541135631907520889568147520
VinculinP18206VCL7176/105.53928313448313964490
Voltage-dependent anion-selective channel protein 1P21796VDAC153173/58.649571860703646181504
X-ray repair cross-complementing protein 5P13010XRCC5785.22/45.695781890310299129272995
X-ray repair cross-complementing protein 6P12956XRCC611511.35/96.23537266058364011947
ZyxinQ15942ZYX804.52/36.23809313748533933499

%Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%.

NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis).

Table 2

All proteins identified in caffeine-exposed EA.hy926 endothelial cells.

Protein nameSwiss-Prot IDGene symbolMS/MS identification score% CovNo. of distinct/total matched peptidespIIntensity (x103 arbitrary unit)
N1N2N3MeanSEM
10 kDa heat shock protein, mitochondrialP61604HSPE1446.91/28.94330388541784131131
14-3-3 protein beta/alphaP31946YWHAB14212.22/54.8201092472721213220161392
14-3-3 protein epsilonP62258YWHAE14213.72/54.65748525852275411169
14-3-3 protein thetaP27348YWHAQ1128.22/44.7295093311131173312641041
14-3-3 protein zeta/deltaP63104YWHAZ14930.66/144.7189252331215327191882309
26S protease regulatory subunit 6BP43686PSMC4326.93/35.1436215282044514469852929
26S protease regulatory subunit 7P35998PSMC24393/45.7138671951313205155282002
26S proteasome non-ATPase regulatory subunit 12O00232PSMD12379.63/77.5239183464333745307693435
26S proteasome non-ATPase regulatory subunit 14O00487PSMD144421.32/36.1NANANANANA
3-ketoacyl-CoA thiolase, mitochondrialP42765ACAA2246.51/18.3NANANANANA
40S ribosomal protein S11P62280RPS113613.32/210.34709698659175871658
40S ribosomal protein S12P25398RPS12109283/66.8182851017813333139322359
40S ribosomal protein S13P62277RPS1312918.53/710.56555388551138511599
40S ribosomal protein S14P62263RPS1412921.23/510.1306013669731621329731885
40S ribosomal protein S15P62841RPS157421.42/310.47814196335966111384281
40S ribosomal protein S16P62249RPS1613114.42/410.2231464158825518300845793
40S ribosomal protein S18P62269RPS186718.42/311.05993605673276458435
40S ribosomal protein S19P39019RPS195912.42/410.39850130011072211191939
40S ribosomal protein S2P15880RPS212817.13/510.3246031569128391228953764
40S ribosomal protein S20P60866RPS208510.11/410.07278763689977971524
40S ribosomal protein S25P62851RPS257881/310.125343395720543781435
40S ribosomal protein S3P23396RPS36910.73/59.74920569358555489288
40S ribosomal protein S30P62861FAU4616.91/212.262664765841764831060
40S ribosomal protein S3aP61247RPS3A4714.83/59.814393120757992114871871
40S ribosomal protein S4, X isoformP62701RPS4X4925.15/710.29693846673398500680
40S ribosomal protein S5P46782RPS57468.82/149.72019383956642125612018846
40S ribosomal protein S6P62753RPS65412.93/410.933367375995168871925
40S ribosomal protein S7P62081RPS79723.74/510.1656095321015787491110
40S ribosomal protein S8P62241RPS8385.31/110.3647310745526474941663
40S ribosomal protein SAP08865RPSA11014.92/34.82129351353123651921
60 kDa heat shock protein, mitochondrialP10809HSPD1104018.37/325.71924234047640473911213180
60S acidic ribosomal protein P0-likeQ8NHW5RPLP0P672184/65.4NANANANANA
60S acidic ribosomal protein P1P05386RPLP133141/14.3436606014935480464307255
60S acidic ribosomal protein P2P05387RPLP25539.12/54.4121111970623420184133328
60S ribosomal protein L10P27635RPL1010812.12/310.1632413597777033254122678
60S ribosomal protein L12P30050RPL1211414.52/39.56275771666776889429
60S ribosomal protein L17P18621RPL173614.13/510.28719654890668111788
60S ribosomal protein L18Q07020RPL184314.42/211.739336316870763191378
60S ribosomal protein L19P84098RPL197013.32/211.57685124607752535890417155
60S ribosomal protein L23P62829RPL2312814.31/310.5552673171001476191304
60S ribosomal protein L23aP62750RPL23A4213.52/210.489901717314570135782414
60S ribosomal protein L27P61353RPL273913.22/310.684723960333252551619
60S ribosomal protein L29P47914RPL29499.41/211.77850486865246414863
60S ribosomal protein L30P62888RPL3093202/39.7102043123411231175566845
60S ribosomal protein L36Q9Y3U8RPL3670192/211.6203072646024430237321810
60S ribosomal protein L38P63173RPL3810735.72/910.1NANANANANA
60S ribosomal protein L4P36578RPL4356.32/311.18221673396808212851
60S ribosomal protein L6Q02878RPL6396.32/310.6191034896549898324639
60S ribosomal protein L7P18124RPL78414.92/410.7NANANANANA
60S ribosomal protein L7aP62424RPL7A2714.73/310.62578298428042788118
78 kDa glucose-regulated proteinP11021HSPA582432.319/395.1NANANANANA
Acidic leucine-rich nuclear phosphoprotein 32 family member AP39687ANP32A1028.41/44.0NANANANANA
Actin, aortic smooth muscleP62736ACTA2203426.812/1985.2NANANANANA
Activator of 90 kDa heat shock protein ATPase homolog 1O95433AHSA1276.51/35.4NANANANANA
Adenylyl cyclase-associated protein 1Q01518CAP13911.23/88.2NANANANANA
ADP/ATP translocase 2P05141SLC25A56815.85/169.740781419793895240571880
ADP-ribosylation factor 1P84077ARF134162/36.3NANANANANA
Alpha-actinin-1P12814ACTN12199.46/95.3NANANANANA
Alpha-actinin-4O43707ACTN434517.212/155.32598931179779629715805651302
Alpha-enolaseP06733ENO1179746.514/2157.016058019570618897618175410764
Ankyrin repeat domain-containing protein SOWAHAQ2M3V2SOWAHA304.42/9710.2833088756565250787086840
Annexin A1P04083ANXA18873910/286.69058013076812149611428112149
Annexin A2P07355ANXA2230651.916/1907.6NANANANANA
Annexin A5P08758ANXA5147435.911/444.924863333940625813328205728805
ATP synthase subunit alpha, mitochondrialP25705ATP5F1A2966.72/79.285448659411111588751513094
ATP synthase subunit beta, mitochondrialP06576ATP5F1B5402710/395.3260942661322905252041159
ATP-dependent RNA helicase DDX1Q92499DDX1995.92/46.8NANANANANA
ATP-dependent RNA helicase DDX3XO00571DDX3X1004.82/46.7NANANANANA
Barrier-to-autointegration factorO75531BANF154271/25.8NANANANANA
Basic leucine zipper and W2 domain-containing protein 1Q7L1Q6BZW11647.42/55.8NANANANANA
Beta-actin-like protein 2Q562R1ACTBL2140317.87/2755.427178410004736148524443976699
CalnexinP27824CANX1032.51/44.5NANANANANA
Calpain small subunit 1P04632CAPNS14691/25.1NANANANANA
Calpain-2 catalytic subunitP17655CAPN2302.11/14.9761512113934796921310
CalreticulinP27797CALR43123/54.34362471445864554103
Casein kinase II subunit alphaP68400CSNK2A1334.61/27.3NANANANANA
Cathepsin DP07339CTSD1568.52/66.1193502698815663206673335
Caveolin-1Q03135CAV111425.33/55.716891907714234134012294
CD59 glycoproteinP13987CD59459.41/36.067791500211515110992383
Cell division control protein 42 homologP60953CDC4216819.93/106.2228623818524789286124819
CentriolinQ7Z7A1CNTRL294.18/145.4NANANANANA
Chloride intracellular channel protein 1O00299CLIC112614.52/45.1352303239031734331181073
Clathrin heavy chain 1Q00610CLTC1114.75/85.5NANANANANA
Cleavage and polyadenylation specificity factor subunit 6Q16630CPSF6738.73/36.7NANANANANA
Coatomer subunit gamma-1Q9Y678COPG1402.52/25.3NANANANANA
Cofilin-1P23528RPS38915.12/38.2170571107711607132471911
Complement component C8 alpha chainP07357C8A412.91/26.189691439811483116171569
Copine-1Q99829CPNE111210.24/265.5598063411528192013042
Core histone macro-H2A.1O75367H2AFY288.12/29.8NANANANANA
Cytoskeleton-associated protein 4Q07065CKAP429720.18/145.6386426475667614570049218
Dehydrogenase/reductase SDR family member 12A0PJE2DHRS12372.51/16.8NANANANANA
Dihydropyrimidinase-related protein 1Q14194CRMP13610.54/56.6363792686346592457510956
DNA damage-binding protein 1Q16531DDB11034.63/55.1NANANANANA
DNA replication licensing factor MCM3P25205MCM3468.95/85.5NANANANANA
DNA replication licensing factor MCM4P33991MCM4305.23/56.3NANANANANA
DNA-(apurinic or apyrimidinic site) lyaseP27695APEX13917.63/48.324785382630747221153
DNA-dependent protein kinase catalytic subunitP78527PRKDC303.411/256.8NANANANANA
Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1P04843RPN1434.42/36.0397564755245760443562357
Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 2P04844RPN2371.91/15.43259178549373327910
E3 ubiquitin/ISG15 ligase TRIM25Q14258TRIM25332.91/18.4NANANANANA
EH domain-containing protein 1Q9H4M9EHD1284.31/16.42850277124562693120
Elongation factor 1-alpha 1P68104EEF1A1181627.17/2009.181440799868321981548935
Elongation factor 1-deltaP29692EEF1D606.82/34.94253385534943868219
Elongation factor 1-gammaP26641EEF1G3611.94/86.320221953715958152393105
Elongation factor 2P13639EEF276626.314/416.415471223610121980220353824862
Elongation factor Tu, mitochondrialP49411TUFM144195/77.322502131221927078214431826
EndoplasminP14625HSP90B133010.27/194.8NANANANANA
Enoyl-CoA hydratase, mitochondrialP30084ECHS11145.91/58.33923425769145032946
Eukaryotic initiation factor 4A-IP60842EIF4A119417.75/95.34303130161188297342735
Eukaryotic translation initiation factor 3 subunit EP60228EIF3E394.31/15.7NANANANANA
Eukaryotic translation initiation factor 3 subunit FO00303EIF3F895.31/55.25340338863095012859
Eukaryotic translation initiation factor 4BP23588EIF4B432.81/25.6NANANANANA
EzrinP15311EZR2929.66/185.9199843995639871332706643
FACT complex subunit SSRP1Q08945SSRP1298.94/56.5NANANANANA
F-actin-capping protein subunit alpha-1P52907CAPZA1426.31/25.5NANANANANA
F-actin-capping protein subunit betaP47756CAPZB8091/15.4NANANANANA
Far upstream element-binding protein 2Q92945KHSRP301.41/16.92971305542573428415
FascinQ16658FSCN1254.91/16.8NANANANANA
Filamin-AP21333FLNA49911.919/285.7NANANANANA
Filamin-BO75369FLNB922.14/65.5417754409239521417961320
Fructose-bisphosphate aldolase AP04075ALDOA51834.99/248.3345703978236191368481540
Galectin-1P09382LGALS138042.25/165.3515175233358343540642152
Gamma-interferon-inducible protein 16Q16666IFI16337.44/59.3NANANANANA
Glucose-6-phosphate isomeraseP06744GPI7710.23/58.4452456709759458572676403
Glucosidase 2 subunit betaP14314PRKCSH587.84/54.3164951920314906168681254
Glutamate dehydrogenase 1, mitochondrialP00367GLUD1748.23/57.7NANANANANA
Glutathione S-transferase PP09211GSTP130341.94/85.410233113861139611005386
Glyceraldehyde-3-phosphate dehydrogenaseP04406GAPDH670954.916/3388.65020874839205043264967786461
Glycerol kinase 2Q14410GK2401.61/55.6NANANANANA
GTP-binding nuclear protein RanP62826RAN8215.73/57.0497614747354665506332121
HEAT repeat-containing protein 5AQ86XA9HEATR5A261.12/1346.1831259226668546813126907
Heat shock 70 kDa protein 1AP0DMV8HSPA1A22410.35/95.5667706178554021608593709
Heat shock 70 kDa protein 6P17066HSPA63089.56/125.8649746090459196616911714
Heat shock cognate 71 kDa proteinP11142HSPA890831.618/375.4NANANANANA
Heat shock protein beta-1P04792HSPB124137.66/126.0437683229127387344824854
Heat shock protein HSP 90-alphaP07900HSP90AA172228.115/404.924362529044026500926635813531
Heat shock protein HSP 90-betaP08238HSP90AB177431.417/385.01836222132452044462004388783
Heterogeneous nuclear ribonucleoprotein D0Q14103HNRNPD6614.44/67.634536167684654891037
Heterogeneous nuclear ribonucleoprotein HP31943HNRNPH125412.54/95.9134401891124674190083243
Heterogeneous nuclear ribonucleoprotein H3P31942HNRNPH3994.91/36.44010312443673834370
Heterogeneous nuclear ribonucleoprotein KP61978HNRNPK13811.24/75.4189191536921312185341726
Heterogeneous nuclear ribonucleoprotein LP14866HNRNPL343.11/18.5NANANANANA
Heterogeneous nuclear ribonucleoprotein MP52272HNRNPM5814.87/98.841711938910860114734404
Heterogeneous nuclear ribonucleoprotein QO60506SYNCRIP647.13/68.7433723364526888346354784
Heterogeneous nuclear ribonucleoprotein RO43390HNRNPR914.12/38.2523070211255182682203
Heterogeneous nuclear ribonucleoprotein UQ00839HNRNPU819.86/95.87785781189598185387
Heterogeneous nuclear ribonucleoproteins A2/B1P22626HNRNPA2B126414.74/89.09234750313746101611861
Heterogeneous nuclear ribonucleoproteins C1/C2P07910HNRNPC745.92/35.0NANANANANA
High mobility group protein HMGI-CP52926HMGA2145333/910.66710715450826315630
Histone H1.4P10412HIST1H1E18721.54/911.02986087832908156950219840
Histone H1.5P16401HIST1H1B3810.62/210.9130582210911606155913286
Histone H2A type 1-B/EP04908HIST1H2AB182428.53/21411.125173825834830171927060215675
Histone H2A type 1-DP20671HIST1H2AD191428.53/21610.9NANANANANA
Histone H2B type 1-BP33778HIST1H2BB797834.94/39510.3NANANANANA
Histone H2B type 1-C/E/F/G/IP62807HIST1H2BC797135.75/40010.3NANANANANA
Histone H2B type 1-MQ99879HIST1H2BM278352.48/9510.3NANANANANA
Histone H3.1tQ16695HIST3H321819.13/1611.1NANANANANA
Histone H4P62805HIST1H4A178051.57/8811.435025530311235901133745917359
Hsc70-interacting proteinP50502ST13305.12/25.23011294211492367609
Importin subunit beta-1Q14974KPNB1303.42/34.7329534167316832543612140
Importin-9Q96P70IPO9683.52/64.7NANANANANA
Integrin beta-4P16144ITGB4443.76/85.74247135051107696092771
Interleukin enhancer-binding factor 2Q12905ILF2278.72/35.212887257799155122123
Interleukin enhancer-binding factor 3Q12906ILF3647.35/58.92673316031733002164
Kelch-like protein 35Q6PF15KLHL35763.92/548.1908229458612395010311910472
Keratin, type II cytoskeletal 7P08729KRT745620.39/255.41105411404041320671276718896
Keratin, type II cytoskeletal 8P05787KRT864333.116/345.51680021801311850181777175058
l-lactate dehydrogenase A chainP00338LDHA38425.98/208.41131221148541228301169362989
l-lactate dehydrogenase B chainP07195LDHB79335.39/225.71513271641731490311548444712
Lysine-specific demethylase 2BQ8NHM5KDM2B315.25/68.9NANANANANA
MoesinP26038MSN76531.217/466.17514310468512199610060813678
Myosin light polypeptide 6P60660MYL614519.22/54.6NANANANANA
Myosin-9P35579MYH960615.123/445.51367221548167516012223324109
Myristoylated alanine-rich C-kinase substrateP29966MARCKS10011.42/34.5NANANANANA
Nascent polypeptide-associated complex subunit alpha, muscle-specific formE9PAV3NACA642.53/39.64236660448315224711
Neurobeachin-like protein 2Q6ZNJ1NBEAL2260.93/56.0NANANANANA
Neuroblast differentiation-associated protein AHNAKQ09666AHNAK1063.914/285.8315402833724712281971972
Nuclear autoantigenic sperm proteinP49321NASP742.91/24.3NANANANANA
NucleolinP19338NCL34012.45/144.6189992996826172250463216
NucleophosminP06748NPM156025.56/224.6702908700371940764115317
Nucleoside diphosphate kinase AP15531NME125628.93/95.8NANANANANA
Nucleoside diphosphate kinase BP22392NME211023.72/58.5NANANANANA
Nucleosome assembly protein 1-like 1P55209NAP1L124411.83/94.4143612247619359187322363
Obg-like ATPase 1Q9NTK5OLA1657.62/37.6118147151373775672341
ParathymosinP20962PTMS4211.81/14.1NANANANANA
Peptidyl-prolyl cis-trans isomerase AP62937PPIA85558.28/447.723170530038723659125622722125
Peptidyl-prolyl cis-trans isomerase BP23284PPIB17528.25/89.4127031891914342153221860
Peroxiredoxin-1Q06830PRDX120648.78/158.3276473011335543311012332
Peroxiredoxin-6P30041PRDX65422.33/66.01613169629042071417
Phenylalanine--tRNA ligase alpha subunitQ9Y285FARSA317.53/47.3116401802114123145951857
Phosphoglycerate kinase 1P00558PGK146830.99/198.3357802923126549305202741
Phosphoglycerate mutase 1P18669PGAM1154223/76.7NANANANANA
Plasminogen activator inhibitor 2P05120SERPINB2406.72/35.51067146264157202421811119
PlectinQ15149PLEC62910.140/765.71295379452611241411215910108
Poly(rC)-binding protein 2Q15366PCBP214310.43/56.3NANANANANA
Polymerase I and transcript release factorQ6NZI2CAVIN12037.72/45.5185692606226052235612496
Polypyrimidine tract-binding protein 1P26599PTBP1778.12/39.2NANANANANA
PRA1 family protein 3O75915ARL6IP57510.11/29.8NANANANANA
Prelamin-A/CP02545LMNA80130.418/426.61783011959631793491845385721
Probable ATP-dependent RNA helicase DDX17Q92841DDX17707.86/118.5100373635721169611852
Probable ATP-dependent RNA helicase DDX5P17844DDX51609.14/69.1608877524568979683704156
Profilin-1P07737PFN117329.34/118.4256823177635538309992871
ProhibitinP35232PHB603.71/35.6356337553652365756
Prohibitin-2Q99623PHB28462/39.8306011607451363932640
Proliferating cell nuclear antigenP12004PCNA8918.83/54.6105211586415787140571768
Proliferation-associated protein 2G4Q9UQ80PA2G46013.54/56.1156064851557986793470
Proteasome activator complex subunit 2Q9UL46PSME2845.41/35.5151067841333772113421
Protein disulfide-isomerase A3P30101PDIA319813.55/96.0144792414214370176643239
Protein disulfide-isomerase A4P13667PDIA4601.91/25.01248301338972719779
Protein disulfide-isomerase A6Q15084PDIA610316.44/75.0158043619531234277446139
Protein MCM10 homologQ7L590MCM10202.62/169.0NANANANANA
Protein S100-A10P60903S100A1015235.12/66.8563653485490549183
Protein S100-A11P31949S100A1197334.33/436.62630494980300015042822301
Protein S100-A6P06703S100A6638.91/25.37360757456556863607
Protein-glutamine gamma-glutamyltransferase 2P21980TGM232016.48/135.1342713610028001327912452
Prothymosin alphaP06454PTMA8712.62/23.71708246519732049222
Proto-oncogene serine/threonine-protein kinase mosP00540MOS512.61/159.2118112864581077131040959315
Purine nucleoside phosphorylaseP00491PNP4712.12/46.54157668552755372731
Putative Ras-related protein Rab-1CQ92928RAB1C3928.93/45.31518215313261666250
Pyruvate dehydrogenase E1 component subunit beta, mitochondrialP11177PDHB558.92/36.2319903211737655339211868
Pyruvate kinase PKMP14618PKM228055.923/1738.039706756786846782447758649547
Rab GDP dissociation inhibitor betaP50395GDI21395.63/86.1401366611371381292895
Ras GTPase-activating-like protein IQGAP1P46940IQGAP1563.74/76.18004678673407376352
Receptor of activated protein C kinase 1P63244RACK14215.54/57.61002333318492061681
Rho GDP-dissociation inhibitor 1P52565ARHGDIA1007.41/25.04867391216513477954
Rho GDP-dissociation inhibitor 2P52566ARHGDIB4812.91/15.1NANANANANA
Rho GTPase-activating protein 26Q9UNA1ARHGAP26252.92/176.2NANANANANA
Ribosome-binding protein 1Q9P2E9RRBP1401.92/38.7NANANANANA
RNA polymerase II-associated protein 1Q9BWH6RPAP1320.71/36.034376928926465431693
RNA-binding protein FUSP35637FUS2318.93/49.4NANANANANA
Serine/threonine-protein phosphatase PP1-gamma catalytic subunitP36873PPP1CC459.92/36.12873562332093902866
Serpin H1P50454SERPINH11258.92/48.8400684792946178447252383
Signal recognition particle 14 kDa proteinP37108SRP1410010.31/110.1NANANANANA
Small nuclear ribonucleoprotein Sm D1P62314SNRPD113816.81/611.6639847268452703297813303
Splicing factor, proline- and glutamine-richP23246SFPQ947.54/109.511573108181360111998831
StathminP16949STMN15415.42/65.8477947520239652642611691
Stress-70 protein, mitochondrialP38646HSPA9478.75/65.9131511938111846147922325
Stress-induced-phosphoprotein 1P31948STIP14515.56/126.447148504941175431439
Sulfotransferase 1A3P0DMM9SULT1A37411.22/45.7NANANANANA
Surfeit locus protein 4O15260SURF4766.71/47.6NANANANANA
T-complex protein 1 subunit alphaP17987TCP1459.94/65.84850104211392197312641
T-complex protein 1 subunit deltaP50991CCT41494.82/38.02671217418722239233
T-complex protein 1 subunit etaQ99832CCT72344.61/97.6NANANANANA
T-complex protein 1 subunit gammaP49368CCT3414.81/16.1NANANANANA
T-complex protein 1 subunit thetaP50990CCT81097.53/55.47786830896448579553
T-complex protein 1 subunit zetaP40227CCT6A9715.65/76.2196612639226299241172228
Thioredoxin domain-containing protein 5Q8NBS9TXNDC5177155/95.655513851143631614211190
TransaldolaseP37837TALDO1413.31/46.415741139478625127712137
Transcription elongation factor A protein-like 3Q969E4TCEAL331243/54.9NANANANANA
Transcription factor BTF3P20290BTF36210.71/29.4NANANANANA
Transgelin-2P37802TAGLN273249.77/288.457399110480767958155815507
Transitional endoplasmic reticulum ATPaseP55072VCP1616.84/125.163742947314137166616787
TransketolaseP29401TKT64311.46/167.6240764735044148385247283
Triosephosphate isomeraseP60174TPI164335.77/225.71462161512481778441584369812
Tropomyosin alpha-1 chainP09493TPM113214.13/84.7NANANANANA
Tropomyosin alpha-3 chainP06753TPM31679.52/64.711546132091304312599529
Tubulin alpha-1A chainQ71U36TUBA1A68831.510/344.9NANANANANA
Tubulin alpha-1B chainP68363TUBA1B70828.49/344.92561152729772684532658485039
Tubulin beta chainP07437TUBB88732.711/354.814636217672114176315494910967
Tubulin beta-4A chainP04350TUBB4A39632.29/174.81213771325441054601197947859
Tubulin beta-4B chainP68371TUBB4B81032.611/334.8NANANANANA
Tubulin beta-6 chainQ9BUF5TUBB618918.86/124.8779418882157231746649265
Ubiquitin carboxyl-terminal hydrolase isozyme L1P09936UCHL18015.22/45.3181811764941400257437830
Ubiquitin-40S ribosomal protein S27aP62979RPS27A37719.93/109.7NANANANANA
Ubiquitin-like modifier-activating enzyme 1P22314UBA133510.77/125.510038413963113681212560912639
UPF0183 protein C16orf70Q9BSU1C16orf70313.61/67.6449612498945433384616738
UPF0258 protein KIAA1024Q9UPX6KIAA1024387.25/87.05248842361916621941
VimentinP08670VIM354055.228/2485.1912676104267293557796364140064
VinculinP18206VCL1576.45/115.5232152573131166267042346
X-ray repair cross-complementing protein 6P12956XRCC619516.46/96.27446319521075105725393
ZyxinQ15942ZYX352.61/16.24116180016542523797

%Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%.

NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis).

Table 3

All proteins identified in EGCG-exposed EA.hy926 endothelial cells.

Protein nameSwiss-Prot IDGene symbolMS/MS identification score% CovNo. of distinct/total matched peptidespIIntensity (x103 arbitrary unit)
N1N2N3MeanSEM
14-3-3 protein beta/alphaP31946YWHAB14212.22/74.8148562576824104215763394
14-3-3 protein epsilonP62258YWHAE12624.34/64.6973711989652394161586
14-3-3 protein thetaP27348YWHAQ7515.54/44.7351663350430882331841247
14-3-3 protein zeta/deltaP63104YWHAZ15045.39/204.7230942875741849312345554
26S protease regulatory subunit 6AP17980PSMC38811.43/55.11779751266425073719010021
26S protease regulatory subunit 6BP43686PSMC4475.33/55.1943618499670360832396984
26S protease regulatory subunit 7P35998PSMC2795.82/45.717425171551488616489805
26S proteasome non-ATPase regulatory subunit 12O00232PSMD12278.33/97.5325633299927461310071778
40S ribosomal protein S11P62280RPS116171/210.32520415054214030839
40S ribosomal protein S12P25398RPS128634.14/76.8109186794375274155233817
40S ribosomal protein S13P62277RPS1315017.92/810.510185826479638804696
40S ribosomal protein S14P62263RPS141037.31/210.11881491672434101384753
40S ribosomal protein S15P62841RPS159413.11/310.416183188124042130124549
40S ribosomal protein S16P62249RPS1617721.93/610.2349802792627421301092440
40S ribosomal protein S18P62269RPS18475.31/211.0546938851001264551836
40S ribosomal protein S19P39019RPS197725.54/410.3416366203441193482886874
40S ribosomal protein S2P15880RPS28419.54/610.3705287870569344728592943
40S ribosomal protein S20P60866RPS206010.11/410.0127861027318891139832559
40S ribosomal protein S24P62847RPS243411.31/310.84519267016272939846
40S ribosomal protein S25P62851RPS258581/210.165222931730555861347
40S ribosomal protein S3P23396RPS34910.72/39.74583657652985485583
40S ribosomal protein S30P62861FAU4916.91/212.2942661561046686831299
40S ribosomal protein S3aP61247RPS3A66113/69.88707724958947283812
40S ribosomal protein S5P46782RPS57177.41/129.7997248287171606847348170
40S ribosomal protein S7P62081RPS711426.85/910.1454811324973285352045
40S ribosomal protein S8P62241RPS86210.62/410.34813613770976016662
40S ribosomal protein SAP08865RPSA1119.52/44.811151877713306110781308
60 kDa heat shock protein, mitochondrialP10809HSPD186919.29/305.7828985862065020688467265
60S acidic ribosomal protein P0-likeQ8NHW5RPLP0P67817.74/55.4NANANANANA
60S acidic ribosomal protein P1P05386RPLP139141/24.3533232116038947378109302
60S acidic ribosomal protein P2P05387RPLP26639.12/34.49083207536118119854467
60S ribosomal protein L10P27635RPL101116.51/310.18641808759334222104
60S ribosomal protein L12P30050RPL1213128.53/59.59704141127536104511935
60S ribosomal protein L15P61313RPL15394.41/111.660401241512841371471
60S ribosomal protein L18Q07020RPL18986.91/411.77596807453527007839
60S ribosomal protein L19P84098RPL19624.61/211.522554190021706153876748
60S ribosomal protein L23P62829RPL239514.31/210.5275565351027065202169
60S ribosomal protein L27P61353RPL27356.61/110.629769151458855721849
60S ribosomal protein L27aP46776RPL27A4911.52/411.0262122703717693236472987
60S ribosomal protein L29P47914RPL29779.41/211.752974889891863681280
60S ribosomal protein L30P62888RPL309513.91/29.727044232358346195425705
60S ribosomal protein L36Q9Y3U8RPL36418.61/111.65046345427483749680
60S ribosomal protein L38P63173RPL389235.72/810.1NANANANANA
60S ribosomal protein L4P36578RPL4432.81/111.11086910149553888521670
60S ribosomal protein L6Q02878RPL6546.32/410.6141766382339979863213
60S ribosomal protein L7P18124RPL710417.34/510.7117027014681685111597
60S ribosomal protein L7aP62424RPL7A7117.75/710.61413391887774103651928
78 kDa glucose-regulated proteinP11021HSPA5108627.816/405.1NANANANANA
Acidic leucine-rich nuclear phosphoprotein 32 family member AP39687ANP32A688.41/44.0NANANANANA
Actin, aortic smooth muscleP62736ACTA2182323.19/1635.2NANANANANA
Activator of 90 kDa heat shock protein ATPase homolog 1O95433AHSA19621.64/55.4NANANANANA
ADP/ATP translocase 2P05141SLC25A56413.14/79.7463701867143459361678788
ADP-ribosylation factor 1P84077ARF1309.91/26.3NANANANANA
Alpha-actinin-1P12814ACTN11499.98/135.3NANANANANA
Alpha-actinin-4O43707ACTN427213.48/115.3902049662181692895064324
Alpha-enolaseP06733ENO1186845.213/4197.020235821299413049618194925909
Annexin A1P04083ANXA1106645.112/356.614878611078410808112255113141
Annexin A2P07355ANXA2165359.319/737.6NANANANANA
Annexin A5P08758ANXA5147541.313/444.947546743388727696439544060441
ATP synthase subunit alpha, mitochondrialP25705ATP5F1A3105.82/69.28346899810108357972117302
ATP synthase subunit beta, mitochondrialP06576ATP5F1B4292710/215.3486373730323580365077244
ATP-dependent RNA helicase AQ08211DHX9323.54/96.41931424531513109668
ATP-dependent RNA helicase DDX3XO00571DDX3X487.43/66.7NANANANANA
Barrier-to-autointegration factorO75531BANF167271/25.8NANANANANA
Basic leucine zipper and W2 domain-containing protein 1Q7L1Q6BZW11718.63/45.8NANANANANA
Beta-actin-like protein 2Q562R1ACTBL2123817.66/2375.435279913598432394327090967975
Beta-enolaseP13929ENO356421.75/807.6NANANANANA
CalnexinP27824CANX1307.13/54.5NANANANANA
Calpain small subunit 1P04632CAPNS15091/25.1NANANANANA
CalreticulinP27797CALR96123/64.328455237812654031527
Cathepsin DP07339CTSD8610.73/66.1334992818524919288672500
Caveolin-1Q03135CAV18713.52/35.7NANANANANA
CD59 glycoproteinP13987CD59289.41/16.0NANANANANA
CentriolinQ7Z7A1CNTRL302.87/135.41090814867324596733411
Chloride intracellular channel protein 1O00299CLIC115212.42/35.1304943286920242278683874
Chloride intracellular channel protein 4Q9Y696CLIC45016.63/65.5NANANANANA
Clathrin heavy chain 1Q00610CLTC1144.55/95.5NANANANANA
Coatomer subunit gamma-1Q9Y678COPG1643.72/25.3NANANANANA
Cofilin-1P23528RPS3786.61/28.2209641653514004171682034
Cofilin-2Q9Y281CFL27011.42/37.7NANANANANA
Copine-1Q99829CPNE113610.24/315.512777914913899119411433
CREB-binding proteinQ92793CREBBP300.51/28.839653520796451501413
Cysteine and glycine-rich protein 1P21291CSRP1408.81/28.9NANANANANA
Cytoplasmic dynein 1 heavy chain 1Q14204DYNC1H1312.810/126.0NANANANANA
Cytoskeleton-associated protein 4Q07065CKAP432825.711/195.6567696640559821609982843
Dedicator of cytokinesis protein 10Q96BY6DOCK102924/106.7285363335318905269324247
Dehydrogenase/reductase SDR family member 12A0PJE2DHRS12382.51/16.8NANANANANA
DNA damage-binding protein 1Q16531DDB11363.43/55.1NANANANANA
DNA-(apurinic or apyrimidinic site) lyaseP27695APEX1545.31/28.36105453546715104502
EH domain-containing protein 1Q9H4M9EHD1427.92/26.44883351934753959462
Elongation factor 1-alpha 1P68104EEF1A1187327.17/3379.1617547713966647685134538
Elongation factor 1-deltaP29692EEF1D11816.74/64.95820611742485395580
Elongation factor 1-gammaP26641EEF1G409.25/76.32145482878522127554350
Elongation factor 2P13639EEF278224.413/346.427381618187011946919171944827
Elongation factor Tu, mitochondrialP49411TUFM10415.34/67.3100052121532940841052238335
EndoplasminP14625HSP90B138410.57/174.8NANANANANA
Enoyl-CoA hydratase, mitochondrialP30084ECHS11265.91/28.31902566694936102104436
Eukaryotic initiation factor 4A-IP60842EIF4A11868.63/75.315563686921492146414246
Eukaryotic translation initiation factor 3 subunit EP60228EIF3E764.31/25.7NANANANANA
Eukaryotic translation initiation factor 3 subunit HO15372EIF3H315.41/16.1986397719072290884
Eukaryotic translation initiation factor 4BP23588EIF4B532.81/25.6NANANANANA
Eukaryotic translation initiation factor 5A-1P63241EIF5A595.21/15.15721375149924821575
Exportin-2P55060CSE1L393.53/2395.516213912437717023715225114132
EzrinP15311EZR29312.18/165.9182752809644421302647625
FACT complex subunit SSRP1Q08945SSRP1656.53/76.5NANANANANA
F-actin-capping protein subunit alpha-1P52907CAPZA1476.31/25.5NANANANANA
F-actin-capping protein subunit betaP47756CAPZB4918.43/65.4NANANANANA
FascinQ16658FSCN1448.53/36.8NANANANANA
F-box/LRR-repeat protein 19Q6PCT2FBXL193022/39.413989149896940119732533
Filamin-AP21333FLNA53412.621/385.712557821212112932215567328244
Filamin-BO75369FLNB1295.510/145.546259120035479827142524310
Fructose-bisphosphate aldolase AP04075ALDOA47734.910/248.3552067681342632582179981
Fructose-bisphosphate aldolase CP09972ALDOC356.31/16.45871376025084046981
Galectin-1P09382LGALS137640.75/145.3535176283355917574222793
Glucose-6-phosphate isomeraseP06744GPI1207.22/58.4115873121211952981107947900
Glucosidase 2 subunit betaP14314PRKCSH7084/64.34119166489265694475011660
Glutathione S-transferase PP09211GSTP131041.94/125.412223117041125911729278
Glyceraldehyde-3-phosphate dehydrogenaseP04406GAPDH649854.614/5648.656647055563252793655001211473
Glycerol kinase 2Q14410GK2491.61/45.6NANANANANA
GTP-binding nuclear protein RanP62826RAN12115.73/67.052227530525504153440835
Heat shock 70 kDa protein 6P17066HSPA63358.95/105.8770967748354634697387553
Heat shock cognate 71 kDa proteinP11142HSPA894626.216/405.4NANANANANA
Heat shock protein beta-1P04792HSPB120438.57/136.01115388927884258950258383
Heat shock protein HSP 90-alphaP07900HSP90AA175620.513/364.925179729892729954628342315814
Heat shock protein HSP 90-betaP08238HSP90AB177125.716/445.019250424204821725121726814302
Hemoglobin subunit alphaP69905HBA16510.61/38.758589805832879971151
Heterogeneous nuclear ribonucleoprotein A1P09651HNRNPA118025.56/89.2271442409023505249131128
Heterogeneous nuclear ribonucleoprotein A3P51991HNRNPA3543.41/29.1NANANANANA
Heterogeneous nuclear ribonucleoprotein D0Q14103HNRNPD3712.43/47.6884778271256897481441
Heterogeneous nuclear ribonucleoprotein HP31943HNRNPH125612.54/115.9269462562423427253331026
Heterogeneous nuclear ribonucleoprotein H3P31942HNRNPH31394.91/26.461096304302351451062
Heterogeneous nuclear ribonucleoprotein KP61978HNRNPK14619.77/105.4386221755617898246926966
Heterogeneous nuclear ribonucleoprotein MP52272HNRNPM5912.37/88.85758214609153121244770
Heterogeneous nuclear ribonucleoprotein QO60506SYNCRIP654.22/68.7137452360321745196983024
Heterogeneous nuclear ribonucleoprotein RO43390HNRNPR764.12/38.291551067486359488612
Heterogeneous nuclear ribonucleoprotein UQ00839HNRNPU1599.96/105.8546110594672975941544
Heterogeneous nuclear ribonucleoproteins A2/B1P22626HNRNPA2B123114.74/199.0134441581431249201695582
Heterogeneous nuclear ribonucleoproteins C1/C2P07910HNRNPC665.92/55.0NANANANANA
Histone H1.0P07305H1F0388.22/210.8548038851001264591835
Histone H1.3P16402HIST1H1D25815.84/1111.0NANANANANA
Histone H1.4P10412HIST1H1E25121.54/811.0308142104147941332657862
Histone H1.5P16401HIST1H1B566.62/310.9410924369938926412391380
Histone H2A type 1-B/EP04908HIST1H2AB80723.12/1411.12264852549582308932374458848
Histone H2A type 1-DP20671HIST1H2AD96223.12/1810.9NANANANANA
Histone H2B type 1-BP33778HIST1H2BB311441.36/52810.3NANANANANA
Histone H2B type 1-C/E/F/G/IP62807HIST1H2BC276435.75/9710.3NANANANANA
Histone H2B type 1-MQ99879HIST1H2BM3154467/56510.3NANANANANA
Histone H3.1tQ16695HIST3H321823.55/1811.1NANANANANA
Histone H4P62805HIST1H4A161855.38/19311.443418239433941744841532311550
Importin subunit beta-1Q14974KPNB1546.14/54.715112133861184313447944
Importin-9Q96P70IPO9282.71/14.7NANANANANA
Inosine-5′-monophosphate dehydrogenase 2P12268IMPDH2564.52/36.4778513828826299581940
Interleukin enhancer-binding factor 3Q12906ILF3745.13/48.93331290753553864755
Kelch-like protein 35Q6PF15KLHL35441.21/328.11245651464908691611932417396
Keratin, type I cytoskeletal 18P05783KRT1846837.411/225.3124666108627930751087899120
Keratin, type II cytoskeletal 1bQ7Z794KRT77396.62/115.7548038851001264591835
Keratin, type II cytoskeletal 7P08729KRT749526.211/275.41191341290761058121180076739
Keratin, type II cytoskeletal 8P05787KRT865136.617/395.51850981870101911851877641797
Laminin subunit beta-1P07942LAMB1353.55/74.8NANANANANA
l-lactate dehydrogenase A chainP00338LDHA34325.98/168.4108915111146977521059384143
l-lactate dehydrogenase B chainP07195LDHB80724.37/215.71700671742721552371665255773
Lysine-specific demethylase 2BQ8NHM5KDM2B391.83/48.9NANANANANA
Macrophage migration inhibitory factorP14174MIF4530.42/37.71123366271121696921532
Malignant T-cell-amplified sequence 1Q9ULC4MCTS1739.41/39.0208021764915260179041605
MoesinP26038MSN91030.816/456.114475210144013868912829413540
Myosin light polypeptide 6P60660MYL615529.13/64.6NANANANANA
Myosin-9P35579MYH949713.217/515.514959518817119792817856514756
Myristoylated alanine-rich C-kinase substrateP29966MARCKS996.91/24.5NANANANANA
Nascent polypeptide-associated complex subunit alpha, muscle-specific formE9PAV3NACA1402.53/59.6196744499544398724908
Neuroblast differentiation-associated protein AHNAKQ09666AHNAK553.718/335.8116441446715574138951170
Non-POU domain-containing octamer-binding proteinQ15233NONO404.91/29.0NANANANANA
Nuclear autoantigenic sperm proteinP49321NASP252.91/14.3NANANANANA
NucleolinP19338NCL33419.99/254.65066335738974107504
NucleophosminP06748NPM163325.56/234.68452748445634596547710465
Nucleoside diphosphate kinase AP15531NME118228.93/145.8NANANANANA
Nucleosome assembly protein 1-like 1P55209NAP1L120892/54.414940179481763116840954
Obg-like ATPase 1Q9NTK5OLA1567.12/37.6133176615578385722385
ParathymosinP20962PTMS8511.81/24.1NANANANANA
Peptidyl-prolyl cis-trans isomerase AP62937PPIA92658.29/467.733490332838227408631245719278
Peptidyl-prolyl cis-trans isomerase BP23284PPIB22722.74/89.4526524318945463471012852
Peroxiredoxin-1Q06830PRDX116749.210/188.3419013756840305399251265
Peroxiredoxin-6P30041PRDX65012.52/36.02427241829252590168
Phosphoglycerate kinase 1P00558PGK136226.98/178.3423604833745011452361729
Phosphoglycerate mutase 1P18669PGAM114717.73/86.7NANANANANA
Plasminogen activator inhibitor 2P05120SERPINB2486.72/35.5286771904312147199564793
Plastin-3P13797PLS310712.45/135.4464295291566457552675900
PlectinQ15149PLEC51211.945/725.733585921202910642921810666300
Poly(rC)-binding protein 2Q15366PCBP213114.24/76.3NANANANANA
Polymerase I and transcript release factorQ6NZI2CAVIN120593/55.5311863704922373302034265
Polypyrimidine tract-binding protein 1P26599PTBP1373.81/29.2NANANANANA
PRA1 family protein 3O75915ARL6IP510214.92/39.8NANANANANA
Prelamin-A/CP02545LMNA75629.416/316.61997682145891952652032075837
Probable ATP-dependent RNA helicase DDX17Q92841DDX17428.25/68.5NANANANANA
Probable ATP-dependent RNA helicase DDX5P17844DDX51024.42/49.1701597262163348687092773
Profilin-1P07737PFN115861.48/138.4525915641450850532851643
ProhibitinP35232PHB323.71/45.67468739046366498931
Prohibitin-2Q99623PHB24041/19.82140450432403295683
Proliferating cell nuclear antigenP12004PCNA4116.93/44.6126592082715557163482391
Proliferation-associated protein 2G4Q9UQ80PA2G4556.32/36.1101991451110920118771333
Proteasome activator complex subunit 2Q9UL46PSME2855.41/25.5117566448784986841588
Protein disulfide-isomerase A3P30101PDIA3160176/96.0235782749824887253211152
Protein disulfide-isomerase A4P13667PDIA4593.42/35.014875548110585103142715
Protein disulfide-isomerase A6Q15084PDIA617616.44/85.0352844372928531358484396
Protein S100-A10P60903S100A1010145.43/66.8NANANANANA
Protein S100-A11P31949S100A11115734.33/416.69892546580676247104315207
Protein S100-A6P06703S100A66516.72/45.314068167317122126412864
Protein transport protein Sec61 subunit betaP60468SEC61B4020.82/211.6NANANANANA
Protein-glutamine gamma-glutamyltransferase 2P21980TGM228016.48/125.1421753924842729413841080
Prothymosin alphaP06454PTMA12512.61/23.7NANANANANA
Proto-oncogene serine/threonine-protein kinase mosP00540MOS426.42/2359.216204412374516967815182214211
Pyruvate dehydrogenase E1 component subunit beta, mitochondrialP11177PDHB687.52/36.2500083828438838423773819
Pyruvate kinase PKMP14618PKM198156.924/1458.054163940557044668746463240291
Rab GDP dissociation inhibitor betaP50395GDI2854.92/76.1148411346510350128861329
Ras-related C3 botulinum toxin substrate 1P63000RAC14515.63/48.8NANANANANA
Ras-related protein Rab-10P61026RAB1084203/48.6NANANANANA
Ras-related protein Rab-1BQ9H0U4RAB1B3721.43/55.6NANANANANA
Ras-related protein Rab-7aP51149RAB7A416.81/26.490071002182299086519
Receptor of activated protein C kinase 1P63244RACK1404.42/37.616698683947166082480
Rho GDP-dissociation inhibitor 1P52565ARHGDIA1137.41/25.072343657441551021088
Ribosome-binding protein 1Q9P2E9RRBP1414.45/68.7NANANANANA
RNA-binding protein FUSP35637FUS2547.82/69.4NANANANANA
Serine/threonine-protein phosphatase PP1-gamma catalytic subunitP36873PPP1CC4814.23/56.157201319515886116003041
Serpin H1P50454SERPINH113914.43/58.882212888339811256389262
Signal recognition particle 14 kDa proteinP37108SRP1417110.31/210.1NANANANANA
Small nuclear ribonucleoprotein Sm D1P62314SNRPD17516.81/511.6366161416513818215337542
Stress-70 protein, mitochondrialP38646HSPA96110.86/95.9424515334545465470873248
Sulfotransferase 1A3P0DMM9SULT1A38311.22/45.7NANANANANA
Surfeit locus protein 4O15260SURF41136.71/47.6NANANANANA
T-complex protein 1 subunit alphaP17987TCP1318.13/35.8NANANANANA
T-complex protein 1 subunit deltaP50991CCT41335.93/48.02861327830143051122
T-complex protein 1 subunit etaQ99832CCT72198.72/97.6NANANANANA
T-complex protein 1 subunit gammaP49368CCT336113/36.1NANANANANA
T-complex protein 1 subunit thetaP50990CCT81075.82/45.4687833851927498464821
T-complex protein 1 subunit zetaP40227CCT6A4711.75/86.2221422278212656191943274
Thioredoxin domain-containing protein 5Q8NBS9TXNDC5206134/65.647384107174781776311681
Tight junction protein ZO-1Q07157TJP1273.75/86.2NANANANANA
TransaldolaseP37837TALDO1314.72/46.4148506899783098602509
Transcription elongation factor A protein-like 3Q969E4TCEAL346202/34.9NANANANANA
Transcription factor BTF3P20290BTF36724.82/39.4NANANANANA
Transgelin-2P37802TAGLN260552.88/268.4965368284276076851516018
Transitional endoplasmic reticulum ATPaseP55072VCP33610.76/175.1481582514619881310628682
TransketolaseP29401TKT611137/227.6624075634945118546245065
Triosephosphate isomeraseP60174TPI162340.68/245.716100618402920848018450513707
Tropomodulin-3Q9NYL9TMOD3569.72/25.13612249235313212361
Tropomyosin alpha-1 chainP09493TPM111615.13/64.7NANANANANA
Tropomyosin alpha-4 chainP67936TPM415618.54/74.7NANANANANA
Tubulin alpha-1B chainP68363TUBA1B73032.810/334.925377528297723545525740213838
Tubulin beta chainP07437TUBB96934.211/364.82028372010221902641980413924
Tubulin beta-4B chainP68371TUBB4B92934.211/344.8NANANANANA
Tubulin beta-6 chainQ9BUF5TUBB622324.28/194.8983919226091928941932101
Ubiquinone biosynthesis monooxygenase COQ6, mitochondrialQ9Y2Z9COQ6351.91/2346.816213912437717023715225114132
Ubiquitin carboxyl-terminal hydrolase isozyme L1P09936UCHL158223/55.313815120839694118641195
Ubiquitin-40S ribosomal protein S27aP62979RPS27A33425.64/139.7NANANANANA
Ubiquitin-like modifier-activating enzyme 1P22314UBA11958.35/125.5947001220971070871079617921
UPF0258 protein KIAA1024Q9UPX6KIAA10243043/47.092365429331959951731
VimentinP08670VIM369359.429/3205.191682596657486124391488030422
VinculinP18206VCL1008.98/125.5223802260710146183784116
Voltage-dependent anion-selective channel protein 1P21796VDAC16517.34/68.618338138943640119574352
X-ray repair cross-complementing protein 6P12956XRCC623213.55/116.212431223657683141604326
ZyxinQ15942ZYX322.11/16.216040880416016136202408

%Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%.

NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis).

All proteins identified in control EA.hy926 endothelial cells. %Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%. NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis). All proteins identified in caffeine-exposed EA.hy926 endothelial cells. %Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%. NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis). All proteins identified in EGCG-exposed EA.hy926 endothelial cells. %Cov = %Sequence coverage = (number of the matched residues/total number of residues in the entire sequence) x 100%. NA = not applicable (protein was identified in the sample but its MS/MS spectra did not meet predefined criteria for high-confident intensity analysis).

Experimental design, materials, and methods

Cell culture

Human endothelial cell line (EA.hy926) (ATCC; Manassas, VA) was grown and maintained in a complete medium (DMEM/F12) (Gibco, Invitrogen; Grand Island, NY), supplemented with 10% heat-inactivated fetal bovine serum (FBS), 60 U/ml penicillin G and 60 μg/ml streptomycin, in a humidified incubator with 5% CO2 at 37 °C.

Treatment with caffeine and EGCG

EA.hy926 cells (from the 5th passage) were seeded in 6-well plate (Corning Inc.; Corning, NY) at a density of 5 × 105 cells/well and maintained in the complete medium for 24-h prior to experiment. The cells were treated with 100 μM caffeine or EGCG (both were from Sigma-Aldrich; St. Louis, MO) for 24-h, whereas the untreated cells served as the control (n = 3 independent biological replicates per group; a total of 9 biological samples were analyzed). Thereafter, the cell monolayers were washed with PBS five times and then scraped. After collecting cell pellets by centrifugation at 1000 g, cellular proteins were extracted using SDT lysis buffer (4% SDS, 100 mM DTT, and 100 mM Tris-HCl; pH 7.6). Protein concentrations were quantified by Bradford's method using Bio-Rad protein assay (Bio-Rad; Milano, Italy).

In-solution tryptic digestion by filter-aided sample preparation (FASP) method

In-solution tryptic digestion was performed as described previously [2], [3]. Briefly, protein samples prepared in SDT lysis buffer were reduced by heating at 95 °C for 5 min. After cooling down at 25 °C, an equal amount of protein (30 μg/sample) was transferred to an Omega Nanosep 10K device (Pall Corporation; Port Washington, NY), added with 200 μl of 8 M urea in 100 mM Tris-HCl (pH 8.5), and then centrifuged at 14,000 g and 25 °C for 15 min. This buffer exchange step was repeated one more cycle. The recovered proteins were then alkylated with 100 μl of 50 mM iodoacetamide in 8 M urea/100 mM Tris-HCl (pH 8.5) at 25 °C in the dark using a ThermoMixer® C (Eppendorf; Hauppauge, NY) for 20 min. Thereafter, buffer exchange was performed twice by centrifugation at 14,000 g and 25 °C for 15 min each using 200 μl of 8 M urea/100 mM Tris-HCl (pH 8.5). The proteins were then finally exchanged into 50 mM NH4HCO3 and digested with sequencing grade modified trypsin (Promega; Madison, WI) in 50 mM NH4HCO3 at a ratio of 1:50 (w/w) trypsin/protein at 37 °C for 16–18 h in a ThermoMixer® C. The digested peptides were collected by transferring the filter unit to a new collection tube and centrifuged at 14,000 g and 25 °C for 10–20 min. Trypsin activity was then stopped by adding 10 μl of 5% formic acid in 80% acetronitrile (ACN), and the digested peptides were dried by a SpeedVac concentrator (Savant; Holbrook, NY). The peptides were finally resuspended in 0.1% formic acid prior to MS/MS analysis.

Analyses of proteins by nanoflow liquid chromatography coupled to tandem mass spectrometry (nanoLC-ESI-Qq-TOF MS/MS)

Separation of the digested peptides was performed using EASY-nLC II (Bruker Daltonics; Bremen, Germany). Briefly, peptides were loaded from a cooled (7 °C) autosampler into an in-house, 3-cm-long pre-column containing 5-μm C18 resin (Dr. Maisch GmbH; Ammerbuch, Germany) and then to an in-house, 10-cm-long analytical column packed with 3-μm C18 resin (Dr. Maisch GmbH) using mobile phase A (0.1% formic acid). The peptides were then separated by mobile phase B (ACN/0.1% formic acid) gradient elution (3–35%) for 150 min at a flow rate of 300 nl/min. Peptide sequences were then analyzed by an ultra-high resolution Qq-TOF MS/MS system (maXis Impact, Bruker Daltonics) in positive mode with ESI nanosprayer ion source. The nanoLC and Qq-TOF MS/MS systems were controlled by HyStar version 3.2 (Bruker Daltonics) and otofControl version 4.1 (Bruker Daltonics), respectively. A capillary voltage and spray shield voltage were set at 5,000V and 500 V, respectively. Nebulizer gas was set at 5.0 psi and dry gas flow rate was at 4.0 l/min, 150 °C [4], [5]. For MS scanning, precursor ions were scanned from 50 to 2200 m/z range (resolution = 40,000 at 622 m/z) and acquired at 2 Hz (0.5 s total accumulation). For MS/MS experiment, the three most intense precursor ions for every MS scan were selected for further fragmentation. Collision-induced dissociation (CID) MS/MS acquisition was performed at 2 Hz (0.5 s total accumulation, if precursor ≤ 1 × 104 ion counts) and 10 Hz (0.1 s total accumulation, if precursor ≥ 5 × 105 ion counts) on the same mass range and resolution set for MS scanning, whereas singly charged ions were excluded. Smart exclusion parameters were set to minimize repeated acquisitions of the same intense precursor ions (repeated count was 2, dynamic exclusion was set at 0.50 min).

MS/MS data processing and protein identification

The raw files (.d) were charge-deconvoluted and extracted into peak list files (.mgf) using Data Analysis version 4.1 software (Bruker Daltonics) via an embedded daMGF script. For identification, the peak list files (.mgf) were searched against the human Swiss-Prot database using Mascot 2.4 search engine (Matrix Science; London, UK). Fixed modification was carbamidomethylation at cysteine residues, whereas variable modification was oxidation at methionine residues. Enzyme was specified to trypsin and only one missed cleavage per peptide was allowed. Data searches were performed with a precursor tolerance of 0.1 Da and fragmentation tolerance of 0.5 Da with +2 and +3 charge state. The false discovery rate (FDR) was performed by searching the decoy database and adjusted to <1% at protein level [5], [6].

Quantitative analysis of MS/MS spectra

Intensity analysis of MS/MS spectra derived from each identified protein was performed using Skyline v.3.5 software (http://proteome.gs.washington.edu/software/skyline). Briefly, The Mascot search result files (.dat) were imported to generate the spectral libraries using BiblioSpec algorithm by the following parameters: spectra cut-off score = 0.95, peptide length = 8–25 amino acids, and share peptides assigned for particular protein. Subsequently, raw files (.d) were then imported and matched to validated peptides in spectral libraries to generate precursor peptide ion intensity. These precursor peptide ion intensities represented the summation of an area under curve of extracted ion chromatograms (XICs) containing three precursor ion isotope peaks (M, M+1, and M+2) [5]. Finally, the total ion intensity of precursor peptide assigned for a given protein was exported and is reported for individual biological replicates, whereas the quantitative intensity data for peptide MS/MS spectra that did not meet the aforementioned predefined criteria are reported as “not applicable”.

Specifications Table

SubjectCell biology
Specific subject areaHuman endothelial cell proteome
Type of dataTable
How data were acquiredMass spectrometry, EASY-nLC II (Bruker Daltonics; Bremen, Germany), Qq-TOF MS/MS system (maXis Impact) (Bruker Daltonics), Mascot 2.4 search engine (Matrix Science; London, UK)
Data formatRaw and Analyzed
Parameters for data collectionEA.hy926 human endothelial cells were treated with 100 μM caffeine or EGCG for 24-h, whereas the untreated cells served as the control. Thereafter, cellular proteins were extracted and subjected to gel-free proteome analysis.
Description of data collectionCellular proteins were subjected to in-solution tryptic digestion and analyzed by nanoLC-ESI-Qq-TOF MS/MS. The mass spectral data were searched against the human Swiss-Prot database using Mascot 2.4 search engine. Fixed modification was carbamidomethylation at cysteine residues, whereas variable modification was oxidation at methionine residues. Enzyme was specified to trypsin and only one missed cleavage per peptide was allowed. Data searches were performed with a precursor tolerance of 0.1 Da and fragmentation tolerance of 0.5 Da with +2 and +3 charge state. The false discovery rate (FDR) was performed by searching the decoy database and adjusted to <1% at protein level.
Data source locationMedical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
Data accessibilityWith the article.
Related research articleAuthor's name: Chanettee Chanthick and Visith ThongboonkerdTitle: Comparative proteomics reveals concordant and discordant biochemical effects of caffeine versus epigallocatechin-3-gallate in human endothelial cells.Journal: Toxicology and Applied Pharmacologyhttps://doi.org/10.1016/j.taap.2019.114621
Value of the Data

These data are the valuable resource for understanding and further analyzing the cellular proteome of human endothelial cells under physiologic condition and after treatment with caffeine or EGCG.

Proteomists, biochemists, physiologists, cellular biologists, and nutritionists will be benefited from these data.

Further functional investigations on these data will lead to better understanding and provide significant insights into vascular cell biology.

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