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XuMass SpectrometryShotgun proteomicshealthy adultsproteomicsAging; healthy adults; plasma; proteomics; tmt-lc-ms/msAgingplasmatmt-lc-ms/mshttps://www.ebi.ac.uk/pride/archive/projects/PXD016199Plasma Cell1. Human plasma collection The plasma samples were collected from the Health Examinations Center of Xinqiao Hospital (The Second Affiliated Hospital), Army Medical University. Samples were collected from 118 adults free of major diseases such as hepatitis, heart disease, infection within six months and immune system-related disease. The participants were divided into three groups: 21-30 (Young), 41-50 (Middle) and ≥60 (Old). Blood (5 ml) was drawn from antecubital vein and collected in ethylenediaminetetraacetic acid (EDTA) tubes, promptly mixed, and centrifuged for 20 min at 1000g at room temperature. Plasma samples were kept in cryotubes at -80℃ until use. 2. Depletion of the plasma and determination of protein concentration Abundant serum proteins were removed from the samples using a Bio-Rad Proteo Miner Protein Enrichment Large-Capacity kit (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions. After that the protein concentration was determined with BCA kit (Beyotime, China) according to the manufacturer’s instructions. 3. Trypsin Digestion The protein solution was mixed with 5 mM dithiothreitol, and kept at 56 °C for 30 min. Then alkylated with 11 mM iodoacetamide for 15 min at room temperature in darkness. After that the protein sample was then diluted by adding 100 mM TEAB to urea concentration less than 2M. Finally, trypsin was added at 1:50 (trypsin:protein) mass ratio for the digestion overnight, then digested with 1:100 (trypsin:protein) mass ratio for 4 h. 4. TMT Labeling After trypsin digestion, peptide was desalted by Strata X C18 SPE column (Phenomenex) and vacuum-dried. Peptide was reconstituted in 0.5 M TEAB, and labeled with TMT kit according to the manufacturer’s instructions. 5. HPLC Fractionation The tryptic peptides were fractionated into fractions by high pH reverse-phase HPLC using Agilent 300Extend C18 column (5 μm particles, 4.6 mm ID, 250 mm length). Briefly, peptides were separated with a gradient of 8% to 32% acetonitrile (pH 9.0) over 60 min into 60 fractions. The peptides were then combined into 18 fractions and dried by vacuum centrifuging. 6. LC-MS/MS Analysis The peptides were dissolved in solvent A (0.1% formic acid), separated by the EASY-nLC 1000 UPLC system on a reversed-phase analytical column (15-cm length, 75 μm i.d.). The gradient setting was as followed: 0~60 min, 6%-20% solvent B (0.1% formic acid in 98% acetonitrile); 60~82 min, 20%-30% solvent B; 82~86 min, 30%-80% solvent B; 86~90 min, 80% solvent B, all at a constant flow rate of 350 nL/min. The peptides were subjected to NSI source followed by tandem mass spectrometry (MS/MS) in Orbitrap fusion lumos (Thermo Fisher). The electrospray voltage applied was 2.4 kV. The m/z scan range was 350 to 1550 for full scan, at a resolution of 60,000. Second scanning started with 100 m/z, at a resolution of 15,000. A data-dependent procedure that alternated between one MS scan followed by 20 MS/MS scans with 15.0s dynamic exclusion. Automatic gain control (AGC) was set at 5E4.PrideThe MS/MS data were processed using Maxquant search engine (v.1.5.2.8). Tandem mass spectra were searched in the Uniprot human database concatenated with reverse decoy database. Trypsin/P was specified as cleavage enzyme, up to 2 missing cleavages were allowed. The mass tolerance for precursor ions in First search and Main search were set as 20 ppm and 5 ppm, respectively, and the mass tolerance for fragment ions was set as 0.02 Da. Carbamidomethyl on Cys was set as fixed modification, and oxidation on Met was set as variable modifications. FDR was adjusted to < 1%. 2. Bioinformatics analysis 2.1 Annotation Methods 2.1.1 Gene Ontology (GO) Annotation Gene Ontology (GO) annotation proteome was derived from the UniProt-GOA database (www. http://www.ebi.ac.uk/GOA/). Briefly, the identified protein ID was converted to UniProt ID and then mapping to GO IDs by protein ID. If some identified proteins were not annotated by UniProt-GOA database, the InterProScan soft would be used to annotated protein’s GO functional based on protein sequence alignment method. Then proteins were classified by Gene Ontology annotation based on three categories: biological process, cellular component and molecular function. 2.1.2 Domain Annotation Identified proteins domain functional description were annotated by InterProScan (a sequence analysis application) based on protein sequence alignment method, and the InterPro domain database (http://www.ebi.ac.uk/interpro/) was used. 2.1.3 KEGG Pathway Annotation Kyoto Encyclopedia of Genes and Genomes (KEGG) database was used to annotate protein pathway. Firstly, using KEGG online service tools KAAS to annotated protein’s KEGG database description. Then mapping the annotation result on the KEGG pathway database using KEGG online service tools KEGG mapper. 2.1.4 Subcellular Localization We used wolfpsort a subcellular localization predication soft to predict subcellular localization. Wolfpsort an updated version of PSORT/PSORT II for the prediction of eukaryotic sequences. 2.2 Functional Enrichment 2.2.1 Enrichment of Gene Ontology analysis Proteins were classified by GO annotation into three categories: biological process, cellular compartment and molecular function. For each category, a two-tailed Fisher’s exact test was employed to test the enrichment of the differentially expressed protein against all identified proteins. The GO with a corrected p-value < 0.05 is considered significant. 2.2.2 Enrichment of pathway analysis KEGG database was used to identify enriched pathways by a two-tailed Fisher’s exact test to test the enrichment of the differentially expressed protein against all identified proteins. The pathway with a corrected p-value < 0.05 was considered significant. These pathways were classified into hierarchical categories according to the KEGG website. 2.2.3 Enrichment of protein domain analysis For each category proteins, InterPro database was researched and a two-tailed Fisher’s exact test was employed to test the enrichment of the differentially expressed protein against all identified proteins. Protein domains with a p-value < 0.05 were considered significant. 2.3 Proteomic phenotyping (Enrichment-based Clustering) Further hierarchical clustering based on different protein functional enrichment was performed as previously reported [16, 17]. We first collated all the categories obtained after enrichment along with their P values, and then filtered for those categories which were at least enriched in one of the clusters with P value < 0.05. This filtered P value matrix was transformed by the function x = −log10 (P value). Finally, these x values were z-transformed for each functional category. These z scores were then clustered by one-way hierarchical clustering (Euclidean distance, average linkage clustering) in Genesis. Cluster membership were visualized by a heat map using the “heatmap.2” function from the “gplots” R-package.ProteomicsXiaoyi XiongDepartment of Neurology, The Second Affiliated Hospital (Xinqiao Hospital), Army Medical University (Third Military Medical University), Chongqing 400037, ChinaPARTIALHomo Sapiens (human)33009535 Xu R, Gong CX, Duan CM, Huang JC, Yang GQ, Yuan JJ, Zhang Q, Xiong XY, Yang QW. Age-Dependent Changes in the Plasma Proteome of Healthy Adults. J Nutr Health Aging. 2020 24(8):846-856 10.1007/s12603-020-1392-6xuruiwade1@sina.comDepartment of Neurology, The Second Affiliated Hospital (Xinqiao Hospital), Army Medical University (Third Military Medical University)China<h4>Objective</h4>Human blood plasma is a complex that communicates with most parts of the body and reflects the changes in the state of an organism. Identifying age-related biomarkers can help predict and monitor age-related physiological decline and diseases and identify new treatments for diseases.<h4>Methods and participants</h4>In this study, TMT-LC-MS/MS was utilized to screen differentially expressed plasma proteins in 118 healthy adults of different ages. Participants were divided into three groups: 21-30 years of age (Young), 41-50 years of age (Middle) and ≥60 years of age (Old).<h4>Results</h4>The number of differentially expressed proteins in the comparisons of Young vs Middle, Middle vs Old and Young vs Old were 82, 22 and 99, respectively. These proteins were involved in numerous physiological processes, such as "negative regulation of smooth muscle cell proliferation" and "blood coagulation". Moreover, when Young was compared with Middle or Old, "complement and coagulation cascades" was the top enriched pathway by KEGG pathway enrichment analysis. Functional phenotyping of the proteome demonstrated that the plasma proteomic profiles of young adults were strikingly dissimilar to those of the middle-aged or older adults.<h4>Conclusions</h4>The results of this study mapped the variation in the expression of plasma proteins and provided information about possible biomarkers/treatments for different age-related functional disorders.Age-Dependent Changes in the Plasma Proteome of Healthy Adults.Xu R R, Gong C X CX, Duan C M CM, Huang J C JC, Yang G Q GQ, Yuan J J JJ, Zhang Q Q, Xiong X Y XY, Yang Q W QWHeart, sodium salt, Dinitrilotetraacetate, ammonium formate, Ethylenedinitrilotetraacetic Acid, 13C-labeled, MeCN, cadmium salt, Tandem, Laboratory, Potassium EDTA, rad, Basodexan, magnesium formate, Vein, zinc salt, Solvent, prevention, Dicobalt, Bca, 5730420M11Rik, Polypeptides, protein polypeptide chains, KL receptor activity, Edetic, cobalt(II) formate dihydrate, Method, CH3-C#N, Heart Disease, B1, High Performance Liquid Chromatography, SCO5, 1, 2, AGM4, SCO1, myd, Gsfsow3, adult, WMS, Sinkiang, (R*, average, Disodium Salt, bca, Fresh Frozen, SET, Divorced, Glycine, N, Mbp-1, proteins, Divorces, W, Edetate, SURGICAL AND MEDICAL PROCEDURES, HPLC, AI047805, DmelCG4299, set, column, venous tree organ part, sample, D1, Chromatography, 2-Ethanediylbis(N-(carboxymethyl)glycine), Bs, s, Homo sapiens disease, High Speed Liquid, Infections and Infestations, Gallium EDTA, plasma, SGS, nickel salt, LC-MS2, Heart Disorder, preventive therapy, subdivided, cobalt (+2) salt, Cardiac Disorder, Dipotassium Salt, Liquid Chromatography, vena, Normalcy, Procedure, heart disorder, ACMICD, PBT, TPSG1, sodium (4:1:1) salt, Dm1, magnesium salt, DmelCG42628, mKIAA0609, Magnesium Salt, University, Raps, Chromium EDTA, forked, fg, Acid, l(3)rF264, pbt, divided, Mainland China, HLA-DR-associated protein II, DI-2, LY57, I-2Dm, formate, dyschromatosis symmetrica hereditaria 1, expanded, Dmel_CG15720, Infection and Infestation, cromium (+3), Methodological, N'-1, septate, beta Trypsin, human, I-2PP1, MDC1D, Sodium Salt, TAF-IBETA, enr, liquid chromatography-tandem mass spectroscopy, PRSS31, ammonium (4:1) salt, TAF-Ibeta, Edetate Disodium Calcium, methyl cyanide, Disodium Versenate, lead (+2) salt, 4-Dimercapto-2, TMT, humans, Ethylenediaminetetraacetic Acid, krk1, Fresh, big, Cleland Reagent, Gallium, disorder of heart/pericardium, rac-Dithiothreitol, nickel formate dihydrate, Fresh Frozen Plasma, lead formate, threo-1, Cleland's reagent, aluminum salt, Ethylenediaminetetraacetic, 4-dimercapto-, Carbamide, R*)-, protein-containing complex, thomson, Normalcies, DmelCG5692, CG7826, Human, mass-to-charge ratio, large, Cardiac, Magnesium Disodium, heart/pericardial disorder, Ly57, LC-MSMS, CG7835, Gene Products, Darknesses, disease or disorder, vascular element, CG42273, 5E4., Carmol, Hepatitides, Man, Technique, Normalities, portion of blood, anatomical systems, pins, thallium (+1) salt, interventionDescription, MDDGB6, heart/pericardial disease or disorder, Monosodium Salt, beta-Trypsin, rubidium salt, 2pp2a, methanoic acid, clinical infection, PINS, High-Performance Liquid Chromatographies, CG10574, Solution, Study, Tetracemate, 2PP2A, Stannous EDTA, Liver inflammation, wvn, dSET, dSet, peptidos, UPLC, Potassium, PTHB1, Drl, 3-butanediol, Individual Health, proto-oncogene c-Kit, CG5692, whole blood, 1-(14)C-labeled, Proteins, disorders, Stannous, Clelands Reagent, Coprin, potassium formate, DSH, copper (+2) salt, backward, acetonitrile, Copper, Dithiothreitol, Mdu, Versene, Frozen Plasmas, polypeptide, Inner Mongolia, Temperatures, native protein, carbonyldiamide, I-2PP2A, mda, KIT ligand receptor activity, C18, cupric formate, chemical analysis, Dm I-2, condition, MFS1, Edathamil, aluminum formate, Pins, ami, DTL, XKrk1, Cleland's, Intervention, AU020952, Heart Disorders, Calcium, DTT, lithium formate, ensemble, med, prophylaxis, Ethylene Dinitrilotetraacetate, Separations, Monopotassium Salt, ethanenitrile, Mass Spectrometry-Mass Spectrometry, CD117, CG10491, Magnesium Disodium EDTA, ME-IV, Cleland's Reagent, Gene Proteins, High Pressure, C-Kit, control, Disodium Ethylene Dinitrilotetraacetate, Ssm, CG4346, xkl-1, liquid chromatography tandem mass spectrometry, Calcium Salt, reversed, l(3)L6A, liquid chromatography tandem mass spectroscopy, IPP2A2, Immune Systems, NCMe, determination, Blood, Mbp1, zinc formate, Infestations and Infections, lead salt, Ethylenedinitrilotetraacetic, protein, Xkl-1, CG15720, Serum, clefted, Calcium Disodium, Mass Spectrometry Mass Spectrometry, heart trouble, CG11121, peptide, Techniques, old, Gsfsco1, ammonium tetraformate, peptido, diseases, Edetic Acid, CT29452, diseases and disorders, Min, protein aggregate, Gsfsco5, prevention and control, Calcium Tetacine, DmelCG42273, SOW3, Fresh Frozen Plasmas, human disease, Edetates, rsh, LCMSMS, Man (Taxonomy), peptides, reference sample, TAF-I, E927b, min, mAPC, Mass Spectrometry, l(3)ddd, copper, SCAN, free, SO, Calcitetracemate, Calcium Disodium Versenate, preventive measures, IGAAD, Immune, lithium salt, High-Performance, Disodium Calcitetracemate, Methodological Studies, Sputolysin, DmelCG10574, High-Performance Liquid, Tetacine, Lyw-57, TEAB, Sl, GPHYSD2, ammonium (2:1) salt, So, Dmel_CG4346, Dm vn, symmetric dyschromatosis of the extremities, phapii, Frozen Plasma, Calcium Disodium Edetate, gyltl1b-b, Modern, LC-MS/MS, Dmel_CG7826, StF-IT-1, Tr-kit, Th, 4-dimercapto-2, Blood Serum, Plasmas, ur, Manchuria, Versenate, SLP65, Ly-57, carbamide, S-adenosyl-L-methionine:thiol S-methyltransferase activity, Cleland, MDDGA6, venae, Distannous EDTA, Diseases, kl1-A, Individual, KIT, KIAA0609, nickel (+2) salt, Adults, Copper EDTA, Dmel_CG7835, Mnb, MNB, tyrosine-protein kinase Kit, SLP-65, ammonium salt, heart/pericardial trouble, Distannous, gyltl1b, BASH, MS/MS, ribosome-associated degradation, System, familial reticulate acropigmentation of Dohi, mdc1d, cobaltous formate, 3-Butanediol, kit, MASS, CG4299, Vn, AW124434, Methodological Study, REM3, R*)-1, disease, H2NC(O)NH2, Health, enlarged, Acetamide, copper salt, DmelCG10491, Liquid, Polypeptide, 2-ethanediylbis(N-(carboxymethyl))-, heart disease or disorder, heart/pericardial disease, RAD1, i2pp2a, 1728, other disease, P1749, human being, Cardiac Diseases, Procedures, Reagent, cesium salt, SCF receptor activity, rapsyn, FBN, Gene, Heart Diseases, Cardiac Disease, froggy, PHAPII, Gyltl1a, LC-MS-MS, disease of heart, formic acid, Intervention or Procedure, Potassium Salt, Karbamid, potassium salt, disease or disorder of heart, Homo sapiens, polypeptide chain, scfr, DmelCG11121, ECTOL1, 14C-labeled, Harnstoff, Studies, SCFR, Separated, Blood Plasma, Fdc, somda, Ethylene, l(3)10568, MS2, l(3)10567, ipp2a2, Interventional, LARGE, labeling, calcium formate, disorder of heart, non-neoplastic, OCTD, BPFD#36, Infestation and Infection, CARDIAC DIS, Chromium Salt, taf-ibeta, EDTA, Blood Plasmas, great, Ultra Performance Liquid Chromatography, disorder, cromium (+3) salt, HEART DIS, Dicobalt EDTA, Disodium EDTA, 2-iodo-, sodium formate, thiol methyltransferase activity, BLNK-S, Controlled, High-Performance Liquid Chromatography, venous subtree, Intervention Strategies, Disease, DYRK1, Controlling, CG42628, nickel formate, 3H-labeled, protein complex, vertebrate blood, igaad, strontium formate, medical condition, strontium salt, Hospital, People's Republic of China, Cardiac Disorders, Peptide, group, Dithiotreitol, LC/MS/MS, venous vessel, natural protein, Systems, Protein, DL-threo-1, I2PP2A, Infection, Dyrk1, Vacuums, Disodium Ethylene, ACETONITRILE, tandem MS, WMS2, cyanomethane, Plasma, Separation, c-KIT, PRSS, Normality, Chelaton 3, uree, Dihydrate, CC1, Tripcellim, portion of plasma, heart disease, Disodium, sample population, chromic formate, Protein Gene Products, Trypure, dSET/TAF-Ibeta, ddd, c-kit, 2610030F17Rik, cardiac disease, l(3)vn10567, concentration, Modern Man, SSKS, calcium salt, Peptid, RAD, Rad, High Performance Liquid, assay, Chromium, AA407739, Serumsliquid chromatography tandem mass spectroscopy, Fresh, other disease, Biological Markers, Viral Marker, human being, multi-cellular organism, Surrogate Endpoints, Fresh Frozen Plasma, Procedures, determination, Clinical Markers, Laboratory, Processes, Clinical Marker, Blood, Clotting, number, Biochemical, Endpoint, Gene, Serum, clefted, LC-MS-MS, CG11121, Human, Surrogate End Points, Physiological Concepts, Surrogate Markers, Laboratory Markers, Techniques, old, Homo sapiens, diseases, DmelCG11121, Biological, Method, LC-MSMS, Studies, Gene Products, disease or disorder, Concepts, diseases and disorders, down-regulation of smooth muscle cell proliferation, animal, Phenomenon, Man, Technique, adult, Biomarker, study, Fresh Frozen Plasmas, Fresh Frozen, Blood Plasma, human disease, somda, LCMSMS, Man (Taxonomy), Clinical, Biological Marker, Physiological Concept, downregulation of smooth muscle cell proliferation, procedures, Blood Clotting, SO, non-neoplastic, Study, Phenomenas, Immunologic Markers, Immune, Markers, Methodological Studies, Viral Markers, Blood Plasmas, Therapies, disorder, Homo sapiens disease, species, Immunologic Marker, plasma, So, Biologic, thiol methyltransferase activity, Physiological, Drl, LC-MS2, Therapy, negative regulation of SMC proliferation, Frozen Plasma, Physiological Process, Viral, Coagulation, subdivided, Surrogate Endpoint, Process, body, RnBP, Modern, Serum Markers, Proteins, disorders, LC-MS/MS, GlcNAc 2-epimerase, End Point, total expressed protein, whole body, Biochemical Markers, medical condition, Procedure, Plasmas, Biologic Marker, results, Immune Marker, Mdu, Concept, Frozen Plasmas, organism, TPSG1, N-acetyl-D-glucosamine 2-epimerase, S-adenosyl-L-methionine:thiol S-methyltransferase activity, LC/MS/MS, inhibition of smooth muscle cell proliferation, Marker, Surrogate End Point, mda, RENBP, Protein, chemical analysis, Diseases, Phenomena, Blood Clottings, condition, down regulation of smooth muscle cell proliferation, techniques, blood clotting, Physiological Phenomenon, Adults, ami, forked, Biologic Markers, non-neoplastic., Plasma, portion of blood plasma, Serum Marker, divided, whole organism, End Points, Surrogate, med, Endpoints, portion of plasma, Immunologic, Methodological, Laboratory Marker, septate, Methodological Study, Treatments, human, Surrogate Marker, AGE, Protein Gene Products, Gene Proteins, disease, blood plasm, Therapeutic, Biochemical Marker, liquid chromatography-tandem mass spectroscopy, Modern Man, Koerper, PRSS31, cardinality, renin-binding protein, liquid chromatography tandem mass spectrometry, Treatment, assay, TMT, Physiological Processes, Proteomes, humans, Elderly, methodology, Immune MarkersPlasma, Fresh, Frozen Plasmas, Fresh Frozen Plasmas, Fresh Frozen, Blood Plasma, N-acetyl-D-glucosamine 2-epimerase, Frozen Plasma, Fresh Frozen Plasma, adult., RnBP, RENBP, Blood Plasmas, Blood, renin-binding protein, GlcNAc 2-epimerase, total expressed protein, portion of plasma, Adults, Plasmas, Proteomes, plasma, AGEliquid chromatography tandem mass spectroscopy, Fresh, Regulations, other disease, Biological Markers, Viral Marker, human being, multi-cellular organism, Surrogate Endpoints, Fresh Frozen Plasma, determination, Clinical Markers, Laboratory, Processes, Clinical Marker, Blood, Myocyte, number, Biochemical, Endpoint, Gene, Smooth Muscle Cells, Serum, clefted, Social Controls, LC-MS-MS, CG11121, Human, Surrogate End Points, Physiological Concepts, Surrogate Markers, Laboratory Markers, Smooth Muscle Cell, old, Homo sapiens, diseases, DmelCG11121, Biological, LC-MSMS, non-striated muscle cell, Gene Products, disease or disorder, Concepts, diseases and disorders, animal, Phenomenon, Man, Formal Social Controls, adult, Biomarker, study, Fresh Frozen Plasmas, Fresh Frozen, Blood Plasma, human disease, somda, LCMSMS, Man (Taxonomy), Clinical, smooth muscle, Smooth Muscle Myocyte, Smooth Muscle, SMCs, myocytes, Biological Marker, Physiological Concept, SO, non-neoplastic, Social, Phenomenas, Immunologic Markers, Immune, Markers, Viral Markers, Blood Plasmas, Therapies, disorder, Homo sapiens disease, species, Smooth Muscle Myocytes, Immunologic Marker, plasma, So, KINDS, Biologic, thiol methyltransferase activity, Physiological, SMC cell, Drl, LC-MS2, Therapy, Frozen Plasma, Physiological Process, Viral, subdivided, Surrogate Endpoint, Process, Formal Social Control, body, RnBP, Modern, Serum Markers, Proteins, disorders, LC-MS/MS, GlcNAc 2-epimerase, End Point, total expressed protein, whole body, Biochemical Markers, medical condition, function, Plasmas, Biologic Marker, Cell, results, Immune Marker, Mdu, Concept, Frozen Plasmas, organism, TPSG1, N-acetyl-D-glucosamine 2-epimerase, S-adenosyl-L-methionine:thiol S-methyltransferase activity, LC/MS/MS, Social Control, Marker, Surrogate End Point, mda, RENBP, Protein, chemical analysis, Diseases, Phenomena, condition, Physiological Phenomenon, Adults, ami, forked, Biologic Markers, non-neoplastic., Plasma, portion of blood plasma, clotting, Serum Marker, divided, whole organism, End Points, Surrogate, med, Endpoints, Control, portion of plasma, Immunologic, Laboratory Marker, Controls, septate, Treatments, human, Surrogate Marker, AGE, Protein Gene Products, Gene Proteins, disease, blood plasm, Therapeutic, Biochemical Marker, liquid chromatography-tandem mass spectroscopy, Modern Man, Koerper, PRSS31, cardinality, Cells, renin-binding protein, liquid chromatography tandem mass spectrometry, Treatment, regulation, assay, TMT, Physiological Processes, Regulation, Proteomes, humans, Immune Markers, smooth muscle fiberIPP2A2, E 920, Gene Ontology Projects, Materials, ion, Hgfr, short stature, single-organism process, determination, methionine aminopeptidase activity, SOFT, Sequence Determination, GIDRP86, cleavage, 5730555F13Rik, protein, soft, 5730420M11Rik, Techniques, protein polypeptide chains, Biological Processes, Biological Phenomenon, caudal, HGFR, Method, Biological, Hot, GIDRP88, Project, AGAMOUS-like 63, Heat, 2, 2-amino-3-mercaptopropanoic acid, Cystein, Galpha47A, establishment and maintenance of substrate location, Analysis, protein aggregate, Phenomenon, WMS, C10orf28, peptidase M activity, CWH3, adenomas, Galphao47A, average, DROGPAD, SET, C, rabGAPLP, L-methionine aminopeptidase activity, Man (Taxonomy), DmelCG2204, HFH2, G-oalpha, Analyses, Genomes, Hfh2, TAF-I, MUB3_18, Determination, cisteina, M, dgo, GORDITA, Sequence Homology Determination, RabGAP-5, proteins, procedures, Goa, GOA, MUB3.18, Sequence Determinations, G(o)alpha47A, Galphao, DmelCG4063, DmelCG4299, IGAAD, set, Methodological Studies, DmelCG10574, RUSC3, ppm, physiological process, onychodysplasia, Tbl1, TBL1, Alignments, RG7MT1, AIS1, GPHYSD2, Racemethionine, Protein Domain, SGS, RNF100, Sequence Homology Determinations, phapii, establishment and maintenance of substance location, proto-oncogene c-Met, Process, PRO, T5E21.12, Modern, 2-Amino-4-(methylthio)butyric acid, MYH-associated polyposis, autosomal recessive familial adenomatous polyposis, StF-IT-1, Maps, single organism process, Search, Peptide Domain, Procedure, Ids, 9130215G10Rik, CYS, Cys, Determinations, ACMICD, single organism localization, Ionen, Domains, Genetic Materials, Methionin, Ontology Projects, cellular component unknown, Hmet, PIX2, Biological Process, (2R)-2-amino-3-sulfanylpropanoic acid, and GLY protein 2, Genetic Material, Domain, T5E21_12, autosomal recessive, parent ion, HGF receptor, HGF/SF receptor, Gene Ontologies, Biologic Phenomena, RUTBC3, Ontology, and GLY protein 1, HLA-DR-associated protein II, Ontologies, CG4063, Projects, function., DI-2, I-2Dm, Sequence Homology, cellular component, MASS, CG4299, Methodological, hCMT1c, c-Met, Methodological Study, E-2f, human, E-2g, L-Cystein, RABGAP5, I-2PP1, goalpha, TAF-IBETA, precursor ion, Material, multiple colorectal, Biocatalyst, facial dysmorphism, Hot Temperatures, Cistron, TAF-Ibeta, i2pp2a, Proteomes, CG2204, humans, cell or subcellular entity, establishment and maintenance of cellular component location, MPS2, human being, 2-amino-4-(methylsulfanyl)butanoic acid, Galpha[[o]], G[[o]]alpha, Go, Procedures, Tb11, Biocatalysts, Processes, L-Zystein, Alignment, FBN, Gene, precursor, dgoalpha, protein-containing complex, DGalpha[[o]], Gene Ontology Project, PHAPII, Gene Ontology, Bkh, Human, FBXW4, and hypotrichosis, method, HGF, Homo sapiens, drogpad, polypeptide chain, ECTOL1, method used in an experiment, Studies, Gene Products, establishment and maintenance of position, DG[[o]], F15E12.6, RCCP2, Man, Technique, familial adenomatous polyposis, MAP, bkh, Search Engines, SIDS, F15E12_6, Galpha[[047A]], Genetic, scatter factor receptor, iones, Ebi, EBI, E-920, familial adenomatous polyposis 2, ipp2a2, 2pp2a, clumped, Hcys, ions, CG10574, AUTS9, G[[o]], OCTD, PSORT, autosomal recessive multiple colorectal adenomas, Study, Enzyme, Peptide Domains, 2PP2A, 2-amino-4-(methylthio)butanoic acid, clustered, 10^[-6], CYSTEINE, taf-ibeta, dSET, dSet, FREE CYSTEINE, AI838057, (2R)-2-amino-3-mercaptopropanoic acid, G[[oalpha]], metionina, molecular function, Biologic, Sequence Analyses, molecular function unknown, DGalpha0, VAMAS2, Galpha[[0]], Encyclopedias, MUTYH-Associated Polyposis, DL-Methionine, protein complex, Proteins, igaad, total expressed protein, tailed, AGL63, F28K20.7, SMAP55, function, Goalpha, Cistrons, Peptide, Engine, group, Sequence Alignments, native protein, Temperatures, natural protein, I-2PP2A, Sequence, Dm I-2, chemical analysis, Protein, I2PP2A, Phenomena, MFS1, establishment and maintenance of localization, techniques, subcellular entity, WDR51A, biological process, Zystein, WMS2, Data Base, ATCMPG1, MET, Met, ATCMPG2, Ontology Project, Temperature, Go-alpha, G-alpha-47A, ensemble, 2-amino-3-sulfanylpropanoic acid, tyrosine-protein kinase Met, L-2-Amino-3-mercaptopropionic acid, (R)-2-amino-3-mercaptopropanoic acid, single-organism localization, MYH-Associated Polyposis, alpha(o), 2-Amino-3-mercaptopropionic acid, l(2)k16213, Protein Gene Products, plan specification, Gene Proteins, dSET/TAF-Ibeta, F28K20_7, 2610030F17Rik, localisation, Ion, Modern Man, SSKS, alpha-amino-gamma-methylmercaptobutyric acid, E920, G-oalpha47a, MAP syndrome, assay, Par4, variable, AA407739, Genesis, 5730455C01Rik, FAP2, SF receptor, methodology, colorectal adenomatous polyposisPlasma, Fresh, Frozen Plasmas, Fresh Frozen Plasmas, Fresh Frozen, Blood Plasma, N-acetyl-D-glucosamine 2-epimerase, Frozen Plasma, Fresh Frozen Plasma, adult., RnBP, RENBP, Blood Plasmas, Blood, renin-binding protein, GlcNAc 2-epimerase, total expressed protein, portion of plasma, Adults, Plasmas, Proteomes, plasma, AGEfalseAge-Dependent Changes in the Plasma Proteome of Healthy AdultsHuman blood plasma is a complex which communicates with most parts of the body and reflects changes in the state of the organism, therefore identifying age-related biomarkers can help to predict and monitor age-related physiological decline and diseases, as well as to find new treatments for diseases. In this study, TMT-LC-MS/MS was utilized to screen for the differentially expressed plasma proteins in 118 healthy adults with different ages. Participants were divided into three groups: 21-30 (Young), 41-50 (Middle) and ≥60 (Old), the number of differentially expressed proteins when Young vs Middle, Middle vs Old and Young vs Old were 82, 22 and 99, respectively. These proteins were found to be involved in numerous physiological processes such as “negative regulation of smooth muscle cell proliferation” and “blood coagulation”. Meanwhile when Young vs Middle or Old, “Complement and coagulation cascades” was confirmed the top enriched pathway by KEGG pathway enrichment analysis. Functional phenotyping of the proteome demonstrated that the plasma proteomic profiles of Young adults were strikingly dissimilar to the Middle or Old adults. The results of this study mapped the variation in expression of plasma proteins, and provided information about possible biomarkers/treatments for different kinds of age-related function disorders.2020-10-192019-11-08PXD016199NEWT:3555NEWT:71647NEWT:330879NEWT:35554NEWT:32046NEWT:544496NEWT:2042546NEWT:32049NEWT:1392696NEWT:259447NEWT:295546NEWT:45351NEWT:445974NEWT:43179NEWT:180454NEWT:5722NCBITaxon:1280NEWT:1129NEWT:309807NEWT:10245NEWT:55153NEWT:309800NEWT:281395NEWT:1211601NEWT:876138NEWT:44271NEWT:193516NEWT:351581NEWT:43186NEWT:428146NEWT:498257NEWT:10036NEWT:1590NEWT:498019NEWT:1351NEWT:1438992NEWT:1352NEWT:2649997NEWT:1147161NEWT:661410NEWT:638632NEWT:263737NEWT:224326NEWT:1333499NCBITaxon:79857NEWT:1096976NEWT:5702NEWT:95648NEWT:1589NEWT:135622NEWT:1349NEWT:35786NEWT:1580NEWT:399784NEWT:96731NEWT:383379NEWT:1401257NEWT:418106NEWT:10029NEWT:913645NEWT:44491NEWT:641809NEWT:27933NEWT:556484NEWT:317447NEWT:4688NEWT:7955NEWT:7719NEWT:7959NEWT:2261NEWT:868565NEWT:31156NEWT:3112NEWT:79329NEWT:4442NEWT:192875NEWT:1214915NEWT:12637NEWT:59729NEWT:2164133NEWT:295105NEWT:108061NEWT:60711NEWT:1316931NEWT:224308NEWT:3347NEWT:511145NEWT:160621NEWT:212790NEWT:931281NEWT:4432NEWT:5762NEWT:5763NEWT:658457NEWT:1310161NEWT:77133NEWT:295358NEWT:145481NCBITaxon:79824NEWT:2246NEWT:2652724NEWT:1912919NEWT:5757NCBITaxon:4563NEWT:5755NEWT:44689NEWT:44447NEWT:752555NEWT:498211NEWT:5759NEWT:347256NEWT:1736231NEWT:5518NEWT:398007NEWT:1527468NEWT:1392NEWT:498217NEWT:498216NEWT:2242NEWT:11320NEWT:286NEWT:391619NEWT:246196NEWT:287NEWT:246197NEWT:1367269NEWT:145458NEWT:633149NEWT:1149786NEWT:10239NEWT:68895NEWT:44685NEWT:161934NEWT:4897NEWT:1148NEWT:5744NEWT:5508NEWT:3329NEWT:5507NEWT:410661NEWT:55571NEWT:35500NEWT:1235816NEWT:1140NCBITaxon:2157NEWT:1143NEWT:4896NEWT:1287689NEWT:1390NEWT:11557NEWT:1094343NEWT:1462472NEWT:1336795NEWT:644042NEWT:294NEWT:1773NEWT:1182590NEWT:3712NEWT:82380NEWT:3711NEWT:935293NEWT:375146NEWT:2065263NEWT:1772NEWT:118698NEWT:1616117NEWT:263NEWT:118696NEWT:52283NEWT:11988NEWT:284812NEWT:8175NEWT:400667NEWT:43330NEWT:1603293NEWT:408169NEWT:44664NEWT:47946NEWT:459758NEWT:3702NEWT:1245466NEWT:243277NEWT:1246791NEWT:7067NEWT:118694NEWT:586722NEWT:2850NEWT:118691NEWT:33548NEWT:274NEWT:7070NCBITaxon:287NEWT:408172NEWT:408170NEWT:96794NEWT:3708NEWT:332648NEWT:44670NEWT:536231NEWT:376219NEWT:219813NEWT:1510NEWT:1436733NEWT:460519NEWT:1247411NEWT:1515NEWT:572307NEWT:1432138NEWT:1424507NEWT:1194599NEWT:272844NEWT:9483NEWT:333760NEWT:56636NEWT:1513458NEWT:1233681NCBITaxon:40559NEWT:506599NEWT:2853422NEWT:81077NEWT:84588NEWT:1679718NEWT:480NEWT:65349NEWT:67767NEWT:46835NEWT:109757NEWT:582580NEWT:300852NEWT:1502NEWT:128017NEWT:376686NEWT:95486NEWT:508771NEWT:1883446NEWT:253NCBITaxon:2759NEWT:1233435NEWT:93061NEWT:93062NEWT:34613NCBITaxon:4932NEWT:109760NEWT:943274NEWT:644223NEWT:235443NEWT:108458NEWT:5936NEWT:272623NEWT:272624NEWT:320637NEWT:886559NEWT:55521NEWT:983964NEWT:118499NEWT:11706NEWT:32644NEWT:527796NEWT:1130798NEWT:55529NEWT:499175NEWT:109779NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:476272NEWT:3747NEWT:195051NEWT:3988NEWT:1116234NEWT:1255228NEWT:649908NEWT:410289NEWT:373153NEWT:3983NEWT:203696NEWT:352472NEWT:1071661NEWT:1202532NEWT:360094NEWT:470NEWT:41364NEWT:5911NEWT:1313NEWT:1218097NCBITaxon:50557NEWT:39251NEWT:39491NEWT:212142NCBITaxon:5811NEWT:29491NEWT:101841NEWT:2014887NEWT:33952NEWT:153009NEWT:261756NEWT:571256NEWT:40483NEWT:63366NEWT:63367NEWT:688333NEWT:215402NCBITaxon:548681NEWT:318586NCBITaxon:5820NEWT:1547NEWT:272634NEWT:198107NEWT:9031NEWT:1872122NEWT:1308NEWT:7091NEWT:1034015NEWT:760568NEWT:141262NEWT:108931NEWT:1055524NEWT:4087NEWT:9778NEWT:306NEWT:150475NEWT:303NEWT:267872NEWT:97477NEWT:7111NEWT:347515NEWT:172269NEWT:9534NEWT:6269NEWT:5180NEWT:256737NEWT:4090NEWT:4092NEWT:9541NEWT:8694NEWT:4093NEWT:4096NEWT:8692NEWT:2903NEWT:185579NEWT:941442NEWT:536088NEWT:13076NEWT:1226408NEWT:7108NEWT:40479NEWT:43989NEWT:4076NEWT:142615NEWT:317NEWT:1006581NEWT:885318NEWT:39488NEWT:550NEWT:4081NEWT:273068NEWT:554NEWT:98334NEWT:451516NEWT:4084NEWT:63577NEWT:36185NEWT:1225786NEWT:7574NEWT:1715256NEWT:30640NEWT:575412NEWT:929793NEWT:29204NEWT:28112NEWT:114155NEWT:6494NEWT:6491NEWT:507601NEWT:520NEWT:186441NEWT:643680NEWT:214092NCBITaxon:6157NEWT:6239NEWT:13095NEWT:162425NEWT:216257NEWT:102169NEWT:208893NEWT:9986NEWT:4054NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:8655NEWT:5147NEWT:98351NEWT:1263854NEWT:118503NEWT:2059687NEWT:160488NEWT:28104NEWT:207559NEWT:407821NCBITaxon:2NEWT:85057NEWT:985076NEWT:568708NEWT:1215323NEWT:986NEWT:52641NEWT:7159NEWT:267671NEWT:28532NEWT:353152NEWT:8496NEWT:7165NEWT:27448NEWT:40674NEWT:1194669NEWT:51329NEWT:243230NEWT:443906NEWT:1080772NEWT:519NEWT:8479NEWT:8485NEWT:6063NEWT:49240NEWT:1328388NEWT:104395NEWT:9557NEWT:6289NEWT:377586NEWT:6287NEWT:300641NEWT:727NEWT:9796NEWT:9554NEWT:118562NEWT:725NEWT:7137NEWT:469008NEWT:256318NEWT:9321NEWT:206411NCBITaxon:6191NEWT:596153NEWT:229533NEWT:1836NEWT:214053NCBITaxon:10298NEWT:80863NEWT:90675NEWT:105231NEWT:104385NEWT:74649NEWT:52638NEWT:57075NEWT:206672NEWT:4097NEWT:8697NEWT:8695NEWT:884204NEWT:71139NEWT:9785NEWT:6279NEWT:1123869NEWT:9544NEWT:9545NEWT:7370NEWT:9793NEWT:83906NCBITaxon:780NCBITaxon:1502NEWT:6282NEWT:1134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