JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST001464/files/F025384-IgG.xlsxhttps://storage.jpostdb.org/JPST001464/files/F025383-IP.xlsxhttps://storage.jpostdb.org/JPST001464/files/Peptide%20Summary%20Report%20(.._data_20200326_F025383.dat).htmlhttps://storage.jpostdb.org/JPST001464/files/Peptide%20Summary%20Report%20(.._data_20200326_F025384.dat).htmlhttps://storage.jpostdb.org/JPST001464/files/MS%20mode.docxprimaryOK200ProteomicsHeng XiaoHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST001464Hunan Normal UniversityjPOSTCo-Immunoprecipitations, keratosis palmaris Et plantaris Familiaris, Co-Immunoprecipitation., EPPK, epidermolytic palmoplantar keratoderma of Voerner, Voerner type, Vörner type, with knuckle pads, CK-9, protein complex, K9, Proteins, VC6rner type, Precipitation, Gene, protein, localized epidermolytic, protein-containing complex, Ppke, palmoplantar keratoderma, Protein Gene Products, Co Immunoprecipitation, Gene Proteins, epidermolytic, Vorner type, epidermolytic palmoplantar keratoderma of VC6rner, tylosis, Immune, native protein, keratoderma, Protein, hyperkeratosis, epidermolytic palmoplantar keratoderma of Vörner, Gene Products, hyperkeratosis palmoplantar localized epidermolytic, Immune Precipitations, Precipitations, protein aggregate, keratosis of Greither, diffuse erythrodermic palmoplantar keratoderma, epidermolytic palmoplantar, Immune PrecipitationIL1BC, Laboratory, CASP-1, Keratin-Associated Proteins, dNSF, mEET, chronic obstructive pulmonary disease (COPD), House Mouse, Polypeptides, T22F8_160, Gamma Globulin, ATGPR7, Chronic irreversible airway obstruction, 1, DGS, GRP8, 2, GRP7, Il1bc, integumentum commune, C79691, (R*, average, Polyacrylamide, SDS-PAGE electrophoresis, N, entire skin, chemical analysis., CAFL - Chronic airflow limitation, Keratin Associated Proteins, IL-1BC, AI047805, Lccp, disease (COPD), sample, Fam91a1, Otf11, Precipitations, AV220772, skin and subcutaneous tissue, NK/GPI, LC-MS2, Gpi, chronic obstructive lung disease [Ambiguous], Caspase-1 subunit p10, Keratin, Malignant Melanomas, antibodies, DmelCG7788, CHRONIC OBSTRUCTIVE AIRWAY DIS, Stickstoff, Dm1, Silver, NK|GPI, Airflow Obstruction, GLYCINE-RICH PROTEIN 8, Chronic Obstructive Airways Disease, Chronic obstructive lung disease, Sodium Dodecyl Sulfate-PAGE, Swiss Mouse, MASCOT, human, CG1106, Speed, nitrogen, B cell receptor accessory molecule complex, F2G1.4, OBSTRUCTIVE PULMONARY DISEASE (COPD), liquid chromatography-tandem mass spectroscopy, GT, Drice, B lymphocyte receptor complex, Mouse, NSF-1, entire integument, Sodium Dodecyl Sulfate-PAGEs, DRICE, hypoderm, 4-Dimercapto-2, Bglap-rs1, Estrogen-enhanced transcript protein, Cleland Reagent, rac-Dithiothreitol, region of skin, Immunoglobulin, Otf-11, threo-1, DrIce, DrICE, Cleland's reagent, Sodium Lauryl, CAO - Chronic airflow obstruction, Nitrate Staining, 4-dimercapto-, NSF1, Gel Electrophoresis, CHRONIC OBSTRUCTIVE PULM DIS, Silver Nitrate Staining, Chronic obstructive pulmonary disease NOS, R*)-, protein-containing complex, SDS polyacrylamide gel electrophoresis of proteins, Chalk, and RNA binding 1, CG7826, Human, Chronic obstructive pulmonary disease finding, melanoma (disease), Electrophoreses, PULMONARY DISEASE (COPD), LC-MSMS, Polyacrylamide Gel Electrophoresis, Immune Precipitations, Gene Products, CG7835, Mus musculus domesticus, CG42273, Pgi, Man, Immune Precipitation, Skn-1a, skin, Immunoglobulin GT, Gpi-1r, Gpi-1s, Naevocarcinoma, Phi, PCE-2, Gpi-1t, SKIN, LITAF-like protein, mKIAA0989, skin organ, Caspase-1 subunit p20, ATGRP7, ATGRP8, NEC, Chronic, peptidos, Associated Protein, SDS PAGE, 3-butanediol, com, Sulfuric acid monododecyl ester sodium salt, Oct-11a, chronic obstructive airways disease NOS, caspase 3, COAD, Proteins, Clelands Reagent, skin zone, CE-2, MF, COLD - Chronic obstructive lung disease, buffer, Amf, chronic, Dithiothreitol, Engine, polypeptide, Sodium Lauryl Sulfate, skin plus hypodermis, Temperatures, native protein, Malignant Melanoma, Sodium, (COPD), chronic airway obstruction, dNSF-1, chronic obstructive, Skin, CAL - Chronic airflow limitation, DTL, NK, skin benign neoplasm, Cleland's, AU020952, DTT, underdeveloped, sodium dodecyl sulphate–polyacrylamide gel electrophoresis, Polyglobin, GR-RBP7, GR-RBP8, House Mice, IL-1 beta-converting enzyme, azote, Interleukin-1 beta-converting enzyme, Epoc-1, ME-IV, Cleland's Reagent, Gene Proteins, chronic obstructive lung disease (disorder), liquid chromatography tandem mass spectrometry, outer epidermal layer, Sulfate, chronic obstructive pulmonary disease, liquid chromatography tandem mass spectroscopy, IgG Antibody, Co-Immunoprecipitations, the integument, Sodium Dodecyl, T22F8.160, Meth, pelt, determination, and rna binding 2, CG 1618, Skn-li, Chronic Airflow Obstructions, Mus domesticus, Dodecyl Sulfate, CG7788, crice, B-cell receptor complex, protein, Co Immunoprecipitation, peptide, peptido, DmNSF, melanoma, chronic obstructive airways disease NOS (disorder), Keratin Associated, SDS-PAGE, dNSF1, dNsf1, Min, Chronic Obstructive Pulmonary Disease (COPD), protein aggregate, Gpi-1, DmelCG42273, COLD (chronic obstructive lung disease), LCMSMS, Man (Taxonomy), peptides, NEC in ICD9CM_2006, membrane bound, Nlk, Co-Immunoprecipitation, dermoid system, Precipitation, hypoplasia, min, Swiss Mice, mAPC, Lauryl Sulfate, outer epithelial layer, LPS-induced TNF-alpha factor homolog, Chronic airflow limitation, alpha-Keratin, Immune, Sputolysin, ICE, Ice, GLYCINE RICH PROTEIN 7, DmelCG1106, house mouse, glycine-rich RNA-binding protein 8, B-lymphocyte receptor complex, CHRONIC OBSTRUCTIVE PULMONARY DISEASE, CHRONIC, obstructive lung disease, iCE, ice, drice, Modern, mouse, LC-MS/MS, Dmel_CG7826, integumental organ, 4-dimercapto-2, Search, Airflow Obstructions, pulmonary disease (COPD), Melanoma, IgG3, mOC-X, IgG4, Cleland, IgG1, drIce, drICE, IgG2, CHRONIC OBSTRUCTIVE, BC033609, chronic obstructive pulmonary disease and allied conditions, Chronic airway disease, NOS, Dmel_CG7835, COPD NOS, Crystal, CHRONIC OBSTRUCTIVE LUNG DIS, Mnb, MNB, dnsf1, Chronic airway obstruction, Mus musculus, Org, ORG, IgG, cold, mice, P45, SDS polyacrylamide gel electrophoresis, Gpi1-r, Gpi1-s, 3-Butanediol, AW124434, Gpi1-t, mKIAA0493, alpha Keratin, domesticus, Chronic Airflow Obstruction, Irium, R*)-1, Silver Nitrate, immunoglobulin, vertebrate epidermis, COPD - Chronic obstructive pulmonary disease, Acetamide, IgG2A, IgG2B, chronic obstructive airways disease, p45, Polypeptide, obstructive pulmonary disease (COPD), 7N, dermal system, chronic obstructive lung disease, COLD, 7S, Gpi1s, Cytokeratin, dermis plus epidermis plus hypodermis, human being, AW549739, AI461847, DISEASE (COPD), Reagent, Comt, Gene, circadian rhythm, Malignant, epidermis, LC-MS-MS, Buffer, NSF, Nsf, antibody, Cold, Sodium Dodecyl Sulfate PAGE, CG1618, Antibody, BCR complex, Pulmonary Disease, Homo sapiens, reduced, House, NEDD4 WW domain-binding protein 3, Tina, tiny, Keratin Associated Protein, Mice, immunoglobulin complex, study, Search Engines, Cold Temperatures, Glass, Staining, IgG(T), Swiss, gel, Chronic Obstructive, DmelCG1618, liquid, epidermis (sensu Metazoa), 2-iodo-, COPD, small, COAD - Chronic obstructive airways disease, DYRK1, bead, protein complex, malignant, not elsewhere classified, hypodermis, PULM DIS CHRONIC OBSTRUCTIVE, Peptide, Chronic obstructive pulmonary disease finding (finding), Dithiotreitol, Melanomas, LC/MS/MS, Oct11, skin region, Mus, Protein, DL-threo-1, Dyrk1, Skin-1a, CES2A1, GLYCINE-RICH RNA-BINDING PROTEIN 7, tegument, Data Base, chronic obstructive airway disease, opsonin activity, Temperature, Dops, CC1, vertebrate integument, sample population, nitrogeno, Laboratory Mice, Interleukin-1 beta convertase, Protein Gene Products, portion of skin, integument, Modern Man, 3.4.22.36, 7S Gamma Globulin, B cell receptor activity, Allerglobuline, malignant melanoma, cold (chronic obstructive lung disease), Peptid, Chronic Obstructive Lung Disease, assay, Laboratory MousefalseIdentifying the hereterodimer partner of KRT9 protein for epidermolytic palmoplantar keratoderma using co-immunoprecipitation coupled with mass spectrometerHuman sole skin sample was collected from resection margins of skin primary malignant melanoma. Most of the sample was stored in liquid nitrogen before being processed for immunoprecipitation. Only epidermis part was used for this study. Skin epidermis was minced and lysed in ice-cold solubilization buffer (P0013, Beyotime). After debris was removed, part of cleared supernatant was transfered to immunoprecipitation using prepared beads bound with mouse monoclonal anti-keratin 9 antibody (ab19124, Abcam) or normal mouse IgG (074-1807, KPL). Proteins that co-precipitated with antibody-bead complex were washed with Tris-buffered saline. Then the beads were resuspended in sample buffer and heated for 5 min at 105℃ to elute proteins. The protein samples were centrifuged and collected for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. Protein bands of interest were excised, reduced with dithiothreitol, alkylated with iodoacetamide, and destained. The gel pieces were digested, and the supernatant was withdrawed for further liquid chromatography-tandem mass spectrometry analysis. Peptide sequences and intensity were analyzed using Thermo Scientific™ Q Exactive mass spectrometer. Protein identification was performed using the Mascot search engine (http://www.matrixscience.com/) by searching LC-MS-MS fragmentation patterns from each peptide against the Swiss-Prot database in Uniprot (https://www.uniprot.org/). Qualified protein sequences that meet the peptide sequencing criterion were retained for afterwards analysis.Sat Jan 28 00:00:00 GMT 2023PXD0313069606