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All rest and exercise plasma samples, along with a representative reference pool (a combination of both haplotypic pairs at rest and during exercise), were processed. Each sample (10 μl), was depleted of albumin and IgG, using the Pierce™ Albumin/IgG Removal Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Subsequently, the modified filter-aided sample preparation method was employed [21]. Half of the volume, corresponding to approximately 30 μg of albumin- and IgG-purified plasma protein, was concentrated on Microcon-30kDa Centrifugal Filter Units (Merck & Co, 865 Kenilworth, NJ, USA), reduced with 100 mM dithiothreitol at room temperature for two hours, washed several times with 8 M urea and once with digestion buffer (DB) (25 mM TEAB, 0.5 % sodium deoxycholate) before alkylation with 10 mM methyl methanethiosulfonate in DB for 30 mins. Digestions were performed by adding 1 μg Pierce mass spectrometry (MS) grade Trypsin (Thermo Fisher Scientific) in DB and incubating overnight at 37°C. Following this, an additional 1 μg trypsin was added, and the samples were incubated for another three hours. Peptides were collected by centrifugation and labeled using Tandem Mass Tag (TMTpro 16plex) reagents (Thermo Fisher Scientific), according to the manufacturer’s instructions. The labeled samples were combined into two sets, where both sets included all four groups and a reference. The paired samples were analyzed in the same set. Sodium deoxycholate was removed by acidification with 10 % TFA and the sets were desalted using Pierce peptide desalting spin columns (Thermo Fisher Scientific) according to the manufacturer’s instructions. The two sets were pre-fractionated into 40 fractions with basic reversed-phase chromatography, using a Dionex Ultimate 3000 UPLC system (Thermo Fisher Scientific), an XBridge BEH C18 column (3.5 μm, 3.0x150 mm, Waters Corp, Milford, MA, USA) and a gradient from 3 % to 90 % acetonitrile in 10 mM ammonium formate with pH 10.0 for 25 mins. The fractions were concatenated into 20 fractions, dried, and reconstituted in 3 % acetonitrile and 0.2 % formic acid.   2.5 Liquid chromatography–Mass Spectrometry/Mass Spectrometry analysis The fractions were analyzed on an Orbitrap Fusion Lumos Tribrid mass spectrometer interfaced with an Easy-nLC 1200 liquid chromatography system (Thermo Fisher Scientific). Peptides were trapped on an Acclaim Pepmap 100 C18 trap column (100 μm x 2 cm, particle size 5 μm, Thermo Fisher Scientific) and separated on an analytical column (75 μm x 35 cm) packed in-house 890 with Reprosil-Pur C18 material (particle size 3 μm, Dr. Maisch, Ammerbuch, Germany) at a flow of 300 nL/min using a gradient from 4 % to 80 % acetonitrile in 0.2 % Formic Acid over 90 mins. The Orbitrap was equipped with the FAIMS Pro ion mobility system alternating between the compensation voltages of -50 and -75. MS scans were performed at 120,000 resolutions, and the 12 most abundant doubly or multiply charged precursors isolated, using a 0.7 m/z isolation window and dynamic exclusion within 10 ppm for 60 seconds. The precursors were fragmented by collision-induced dissociation (35 %) and detected in the ion trap, followed by multi-notch (simultaneous) isolation of the top 10 MS2 fragment ions within the m/z range 400-1400, MS3 fragmentation by higher-energy collision dissociation (55 %) and detection of the reporter ions in the Orbitrap at 50 000 resolution, m/z range 100-500."],"repository":["Pride"],"modification":[""],"quantification_method":[""],"data_protocol":["2.6 Proteomic data analysis The data files in each set were merged for identification and relative quantification using Proteome Discoverer version 2.4 (Thermo Fisher Scientific). The Uniprot Equus caballus proteome database (July 2021, combined SwissProt and TrEMBL 44,488 entries) and Mascot search engine v. 2.5.1 (Matrix Science, London, UK) were used for database matching with peptide and fragment ion tolerance of 5 ppm and 0.6 Da. Tryptic peptides with zero missed cleavage, variable modifications of methionine oxidation and fixed cysteine alkylation were accepted. ProTMT modifications of N-terminal and lysine were selected. A percolator was used for the peptide-spectrum match validation with a strict false discovery rate (FDR) threshold of 1 %. The TMT reporter ions were identified with 3 mmu mass tolerance and no normalization was applied to the samples. For the protein quantification, only unique peptide sequences at a strict FDR threshold of 1 % with a minimum synchronous precursor selection match of 65 % and an average signal:noise ratio above 10, were taken into account. The reference in each set was used to calculate protein ratios, and quantified proteins were filtered at 5 % FDR."],"omics_type":["Proteomics"],"labhead":["Gabriella Lindgren"],"instrument_platform":[""],"labhead_affiliation":["Department of Animal Breeding and Genetics Swedish University of Agricultural Sciences"],"submission_type":["PARTIAL"],"species":["Equus Caballus (horse)","Homo Sapiens (human)"],"publication":["38906044 Amiri Roudbar M, Rosengren MK, Mousavi SF, Fegraeus K, Naboulsi R, Meadows JRS, Lindgren G. Effect of an endothelial regulatory module on plasma proteomics in exercising horses. Comp Biochem Physiol Part D Genomics Proteomics. 2024 52:101265 10.1016/j.cbd.2024.101265"],"submitter_mail":["johannes.fuchs@gu.se"],"submitter_affiliation":["Gothenburg University"],"submitter_country":["Sweden"],"pubmed_abstract":["Elite performing exercise requires an intricate modulation of the blood pressure to support the working muscles with oxygen. We have previously identified a genomic regulatory module that associates with differences in blood pressures of importance for elite performance in racehorses. This study aimed to determine the effect of the regulatory module on the protein repertoire. We sampled plasma from 12 Coldblooded trotters divided into two endothelial regulatory module haplotype groups, a sub-elite performing haplotype (SPH) and an elite performing haplotype (EPH), each at rest and exercise. The haplotype groups and their interaction were interrogated in two analyses, i) individual paired ratio analysis for identifying differentially abundant proteins of exercise (DAPE) and interaction (DAPI) between haplotype and exercise, and ii) unpaired ratio analysis for identifying differentially abundant protein of haplotype (DAPH). The proteomics analyses revealed a widespread change in plasma protein content during exercise, with a decreased tendency in protein abundance that is mainly related to lung function, tissue fluids, metabolism, calcium ion pathway and cellular energy metabolism. Furthermore, we provide the first investigation of the proteome variation due to the interaction between exercise and related blood pressure haplotypes, which this difference was related to a faster switch to the lipoprotein and lipid metabolism during exercise for EPH. The molecular signatures identified in the present study contribute to an improved understanding of exercise-related blood pressure regulation."],"pubmed_title":["Effect of an endothelial regulatory module on plasma proteomics in exercising horses."],"pubmed_authors":["Amiri Roudbar Mahmoud M, Rosengren Maria K MK, Mousavi Seyedeh Fatemeh SF, Fegraeus Kim K, Naboulsi Rakan R, Meadows Jennifer R S JRS, Lindgren Gabriella G"],"additional_accession":[]},"is_claimable":false,"name":"Effect of an endothelial regulatory module on plasma proteomics in exercising horses","description":"Elite performing exercise requires an intricate modulation of the blood pressure to support the working muscles with oxygen. We have previously identified a genomic regulatory module that associates with differences in blood pressures of importance for elite performance in racehorses. This study aimed to determine the effect of the regulatory module on the protein repertoire. We sampled plasma from 12 Coldblooded trotters divided into two endothelial regulatory module haplotype groups, a sub-elite performing haplotype (SPH) and an elite performing haplotype (EPH). By sampling each at rest and exercise, we aimed to quantify the regulatory module's effect on blood pressure. The haplotype groups and their interaction were interrogated in two analyses, i) individual paired ratio analysis for identifying differentially abundant proteins of exercise (DAPE) and interaction (DAPI) between haplotype and exercise, and ii) unpaired ratio analysis for identifying differentially abundant protein of haplotype (DAPH). At an expression fold change >1.5, our analyses identified 39 (DAPE), 53 (DAPH) and 20 (DAPI) proteins that showed significantly different expression levels (p-value ≤ 0.05). The proteomics analyses revealed a widespread change in plasma protein content during exercise, regardless of haplotype status, with a decrease tendency in protein abundance that mainly related to lung function, tissue fluids, metabolism, calcium ion pathway and cellular energy metabolism. Furthermore, we provide the first investigation of the proteome variation due to interaction between exercise and related blood pressure haplotypes, which this difference was related to a faster switch to the lipoprotein and lipid metabolism during exercise for EPH. The molecular signatures identified in the present study contribute to improved understanding of the biological mechanisms behind exercise and exercise-related blood pressure regulation of the module that affect racing performance in horses.","dates":{"publication":"2026-07-06","submission":"2023-12-14"},"accession":"PXD047817","cross_references":{"TAXONOMY":["NEWT:330879","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:112509","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:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:1907535","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","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: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:1912919","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:37919","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:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","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: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:8165","NEWT:96794","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","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:56636","NEWT:2709072","NEWT:2499525","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:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","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: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:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","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:30455","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:299467","NEWT:9541","NEWT:51350","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","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