{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Msf":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/melanocytes.msf"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/checksum.txt"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/14a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/9a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/17a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/22a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/11a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/2a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/18a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/8a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/5a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/23a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/12a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/15a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/4a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/19a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/7a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/20a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/24a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/16a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/21a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/10a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/13a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/6a.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074276/3a.raw"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["agnieszkagegotek@o2.pl"],"submitter":["Agnieszka Gęgotek"],"technology_type":["Mass Spectrometry","Gel-based experiment"],"software":[""],"submitter_keywords":["proteomics","3-o-ethyl ascorbic acid","Melanocytes","Melanocytes; uva radiation; cannabigerol; 3-o-ethyl ascorbic acid; proteomics; skin protection;","cannabigerol","skin protection","uva radiation"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD074276"],"tissue":["Melanocyte","Cell Culture"],"sample_protocol":["Cells culture and treatment Human primary epidermal melanocytes (ATCC PCS-200-012) isolated from the foreskin of a newborn were obtained from the American Type Culture Collection ATCC® (Manassas, VA, USA). Cells were cultured according to the manufacturer guidelines in a humid atmosphere of 5% CO2 and a temperature of 37 °C, using Dermal Cell Basal Medium (DCBM, ATCC-PCS-200-030) supplemented with Melanocyte Growth Kit (ATCC-PCS-200-041) and antibiotics: 50 U/mL penicillin and 50 µg/mL streptomycin. Melanocytes from passage 3 were used in the experiment.  After reaching 90% confluency, cells were exposed to UVA radiation with a total dose 18 J/cm2 (365 nm; intensity of 4.08 mW/cm²; Bio-Link Crosslinker BLX 365; Vilber Lourmat, Germany). The radiation dose was chosen in reference to approximately 70% of cell viability as measured by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method [18]. To examine the effect of CBG and EAA, melanocytes were incubated for 24 hours in a medium supplemented with CBG (0.5-15 µM; obtained after appropriate dilution from a 32 mM solution of CBG in ethanol; final ethanol concentration in medium was 0.3%) and/or EAA (50-250 µM, diluted in phosphate-buffered saline, PBS). Independently, untreated and unirradiated cells were cultured as a control group. To eliminate the ethanol’s differentiating effect on the cells, all groups were cultured in a medium containing 0.3% of ethanol. After 24 h incubation, a viability test and proteomic analyses were performed. Proteomic analysis Approximately 2x106 cells per plate following UVA irradiation and/or treatment with CBG/EAA were scraped to cold PBS and disintegrated by sonification on ice. Following centrifugation, the total protein content of the cell lysate was measured using the Bradford assay [19]. Volume of lysates containing 30 µg were electrophoretically separated on 10% SDS-PAGE gels. Gels were stained with Coomassie brilliant blue, and complete sample lanes were cut out. Proteins in gel parts were reduced with 10 mM dithiothreitol and alkylated with 50 mM iodoacetamide. Following overnight in-gel digestion using trypsin (Promega, Madison, WI, USA), obtained peptide mixtures were dried under inert gas and dissolved in 5% acetonitrile (with 0.1% formic acid). Direct peptide sequence analysis was performed using a QExactive HF mass spectrometer with an electrospray ionization source operated using Xcalibur 4.1 (Thermo Fisher Scientific, Bremen, Germany) following preliminary peptides separation on the 150 mm x 75 µm PepMap RSLC capillary analytical C18 column (Dionex, LC Packings) using Ultimate 3000 (Dionex, Idstein, Germany) as described previously [20]. The mass spectrometer was operated in positive mode and survey MS scans were conducted in the 200–2000 m/z range with a resolution of 120,000. In subsequent scans, the top ten most intense ions were isolated, fragmented, and analyzed at 30,000 resolution. Differences between samples in the intensity of randomly selected peaks were validated using an Agilent 1290 Infinity II LC system coupled to an Agilent 6475 triple quadrupole LC/MS system (Agilent Technologies, Heilbronn, Germany)."],"repository":["pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Obtained raw data were processed using Proteome Discoverer 2.0 (Thermo Fisher Scientific, Bremen, Germany), and input data were searched against the UniProtKB - SwissProt database (taxonomy: Homo sapiens, release 2025-04). As a dynamic modification of the chosen amino acids (cysteine, lysine, and histidine), adducts formation with 4-HNE was set [21]. Only proteins with at least two unique peptides longer than 6 amino acid residues were taken for statistical analysis. The signal intensities of the precursor ions were used for label-free quantification of proteins.  The results from proteomics, including individual protein label-free quantification were subjected to data imputation (missing values were replaced by 1/5 of the minimal positive values of their corresponding variables), normalized by the median of the protein intensities, and log-transformed using open-source software MetaboAnalyst 6.0 (http://www.metaboanalyst.ca; accessed on 11 September 2025) [22]."],"omics_type":["Proteomics"],"labhead":["Agnieszka Gęgotek"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["UMB"],"species":["Homo Sapiens (human)"],"submitter_mail":["agnieszkagegotek@o2.pl"],"publication":["42274558 Mucha M, Ryšavá A, Jarocka-Karpowicz I, Maruška A, Skrzydlewska E, Gęgotek A. Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-&lt;i&gt;O&lt;/i&gt;-Ethylascorbic Acid. Cells. 2026 15(11):965 10.3390/cells15110965"],"submitter_affiliation":["Medical University of Białystok"],"submitter_country":["Poland"],"pubmed_abstract":["The aim of this study was to analyze the effect of concomitant use of cannabigerol (CBG) and 3-<i>O</i>-ethylascorbic acid (EAA) on changes in the proteome of UVA-irradiated skin melanocytes, with particular emphasis on adduct formation between lipid peroxidation products and metabolically important proteins. Proteomic analysis allowed the identification of 1248 proteins with statistically significantly changed expression following melanocytes irradiation and/or incubation with CBG/EAA. The top 25 proteins with the most strongly differentially abundant expression included proteins involved in cell protection/antioxidant response, as well as pro-inflammatory and proapoptotic signalization. Moreover, in melanocytes irradiated with UVA, the levels of lipid peroxidation product, 4-hydroxynonenal (4-HNE) and its protein adducts were increased, as well as significant changes in the profile of proteins modified by 4-HNE were observed. CBG and EAA, especially when used together, largely reverse these effects. This study for the first time demonstrated the combined effect of CBG and EAA on the proteome of melanocytes after their exposure to UVA radiation, which applies to both changes in protein expression and intracellular signaling based on proteins modified by 4-HNE. It can be suggested that CBG and EAA may provide melanocytes with effective protection against the effects of oxidative stress and perhaps even protect the skin from carcinogenesis."],"pubmed_title":["Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-&lt;i&gt;O&lt;/i&gt;-Ethylascorbic Acid."],"pubmed_authors":["Mucha Magda M, Ryšavá Alena A, Jarocka-Karpowicz Iwona I, Maruška Audrius A, Skrzydlewska Elżbieta E, Gęgotek Agnieszka A"],"additional_accession":[]},"is_claimable":false,"name":"CBG and EAA for melanocytes protection","description":"Aims: This study aims to assess the effect of simultaneous use of cannabigerol (CBG) and 3-O-ethyl ascorbic acid (EAA) on changes in the skin melanocyte proteome induced by UVA radiation, with particular emphasis on modifications in the structure of metabolically important proteins. Results: Proteomic analysis allowed to identify 1248 proteins with statistically significantly changed expression following melanocytes irradiation and/or incubation with CBG/EAA. The top 25 proteins with most strongly modified expression included mainly proteins involved in cells protection/antioxidant response, as well as pro-inflammatory and proapoptotic signalization. Moreover, in melanocytes irradiated with UVA, the levels of lipid peroxidation product, 4-hydroxynonenal (4-HNE) and its protein adducts increase, as well as significant changes in the profile of proteins modified by 4-HNE are observed. CBG and EAA, especially when used together, largely reverse this effect. Innovation: This study for the first time shows the combined effect of CBG and EAA on the proteome of melanocytes after their exposure to UVA radiation, that applies to both changes in protein expression and intracellular signaling based on protein modification by lipid peroxidation products. Conclusion: It can be summarized that CBG and EAA can provide melanocytes with the most effective protection against oxidative stress, and perhaps even protect the skin structure against carcinogenesis.","dates":{"publication":"2026-06-15","submission":"2026-02-10"},"accession":"PXD074276","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","NEWT:475174","NEWT:111205","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:111225","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:342808","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:156981","NEWT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