{"database":"MassIVE","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://massive-ftp.ucsd.edu/v07/MSV000096340/"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"submitter":["Malgorzata Przybylo"],"full_dataset_link":["https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=2d3eb396f7df48a7955071484077ef2d"],"submitter_email":["malgorzata.przybylo@uj.edu.pl"],"sample_protocol":[""],"repository":["MassIVE"],"file_size":["28"],"ptm_modification":["UNIMOD:4 - \"Iodoacetamide derivative.\"","UNIMOD:1 - \"Acetylation.\"","UNIMOD:35 - \"Oxidation or Hydroxylation.\""],"data_protocol":[""],"omics_type":["Proteomics"],"instrument_platform":["Q Exactive"],"species":["Homo Sapiens (ncbitaxon:9606)"],"submitter_affiliation":["Department of Glycoconjugate Biochemistry, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University"],"pubmed_abstract":["Extracellular vesicles (EVs) are potential non-invasive diagnostic, prognostic and therapeutic tools. Additionally, they are important contributors to tumorigenesis. Glycosylation has been found to modulate the composition of the EV proteome. Increased amounts of β1,6-branched N-glycans, synthesized by N-acetylglucosaminyltransferase V (GnT-V), are most commonly observed in melanoma and are associated with decreased cell adhesion and increased metastasis. The opposite effect is caused by the addition of bisecting GlcNAc by N-acetylglucosaminyltransferase III (GnT-III). To date, the impact of these enzymes on EV cargo in melanoma remains unexplored. Flow cytometry was used to study the surface glycosylation of genetic variants of WM266-4 melanoma cells with induced overexpression of GnT-III or GnT-V encoding genes (MGAT3 or MGAT5) and EVs released by these cells. LC-MS/MS proteomics was applied to analyze the effect of altered glycosylation on the proteome of released EVs, followed by detailed bioinformatic analysis. Flow cytometry analysis revealed dynamic changes in the surface glycosylation of EVs derived from melanoma cells overexpressing MGAT3 or MGAT5. Induced overexpression of MGAT3 or MGAT5 also caused significant changes in the proteome of EVs. The proteomic analysis identified a total of 1770 microvesicular and 704 exosomal proteins that play different roles in melanoma progression, including those with established diagnostic/prognostic potential and those closely associated with melanoma onset. Proteomic profiling of EVs derived from cells overexpressing MGAT3 and MGAT5 revealed functional changes in EV protein content driven by glycosylation modifications. The study presented a potential multifaced application of melanoma-derived EVs for diagnostic and prognostic purposes."],"pubmed_title":["MGAT3 and MGAT5 overexpression alters the protein cargo of extracellular vesicles released by metastatic melanoma cells."],"pubmed_authors":["Wilczak Magdalena M, Surman Magdalena M, Jankowska Urszula U, Skupien-Rabian Bozena B, Przybyło Małgorzata M"],"pubmed_abstract_synonyms":["GNT-III protein, liquid chromatography tandem mass spectroscopy, Glc-N-Ac-transferase V, Materials, O-linked Glycosylations, determination, Metastasis, 6 N-acetylglucosaminyltransferase V, Galactosylation, Galactosylations, Glykan, Neoplasm Metastases, Extracellular Vesicle, protein, neoplasm metastasis, cargo, Molecular Biologies, cancer metastasis, polisacarido, rat, protein polypeptide chains, Bio Informatics, 6)N-acetylglucosaminyltransferase, melanoma, GnT-III protein, Roles, Concepts, 3, ramiform, Flow Microfluorimetry, protein aggregate, Cell Sortings, integumentum commune, imprinted and ancient gene protein, treatment, LCMSMS, enzymes, Glycans, Fluorescence Activated Cell Sorting, Biology, A330041C17Rik, N-Acetylglucosaminylation, Glycane, Age of onset, Oncogeneses, proteins, ramified, Bio-Informatic, Sialylation, Computational, alpha-1, genetic, Adhesions, Bio-Informatics, O-linked, gnt, Adhesion, Glycan, Role Concepts, disease management, Therapies, CG7776, Computational Molecular Biologies, N-Acetylglucosaminylations, Flow Cytofluorometry, Mgat5 protein, LC-MS2, Therapy, Cell Sorting, N-glycosyl-oligosaccharide-glycoprotein N-acetylglucosaminyltransferase V, Fluorescence-Activated Cell Sorting, external covering of organism, Biologies, mouse, e(Pc), familial, LC-MS/MS, beta-1, Glycosylations, organism surface, Malignant Melanomas, L[[3]]CP3, Fucosylations, Computational Molecular, Glykane, Microfluorimetry, Melanoma, Role Concept, UDP-N-acetylglucosamine alpha-6-D-mannoside beta1, mannosyl (beta-1, Phosphoglycosylations, Playthings and Play, Apoptotic, Computational Molecular Biology, Role, Plaything, Genetic Materials, Genetic Material, Cytofluorometries, DmelCG7776, Bioinformatic, N linked Glycosylation, Toys, Sialylations, Fluorescence-Activated Cell, Treatments, human, MGAT3 protein, polysaccharides, GnT V enzyme, 4-)-glycoprotein beta-1, Mgat3 protein, Oncogenesis, integumentary system, Material, N Acetylglucosaminylation, liquid chromatography-tandem mass spectroscopy, DmelLcp3, E(PC), BG:DS02740.11, anon-48Ac, Biocatalyst, Cistron, inherited genetic, E430016J11Rik, dermal system, Proteomes, 4-N-acetylglucosaminyltransferase III, GH05739, Tumorigeneses, Plays, Exovesicles, GlcNAcylation, Apoptotic Body, Bodies, Peptidomics, Biocatalysts, Vesicles, glycans, Gene, N-linked Glycosylations, protein-containing complex, Malignant, Ximpact, LC-MS-MS, N-linked, Fluorescence-Activated, melanoma (disease), surface, Fluorescence-Activated Cell Sortings, Microfluorometry, O-linked Glycosylation, mannoside acetylglucosaminyltransferase 3, III, glycosylation, N-acetylglucosaminyltransferase V, GH14582, polypeptide chain, Glycosylation, 3(6)-mannosylglycoprotein beta-1, LC-MSMS, Gene Products, single organism cell adhesion, Polysaccharide, Cytofluorometry, Toy, DmelCG2043, study, tumor cell migration, lcp3, Genetic, Playthings, Flow Cytofluorometries, Protein Glycosylations, cell adhesion molecule activity, Microfluorometries, Naevocarcinoma, LCP-3, metastasis, mannoside acetyl glucosaminyltransferase 3, Enzyme, CP3, Puppets, Phosphoglycosylation, 6-N-acetylglucosaminyltransferase, Extracellular, Flow, Flow Microfluorometries, Cytometry, Play, Fucosylation, constitutitional genetic, tumor metastasis, UDPGNAC-MAGtransferase III, N-linked Glycosylation, Puppet, Cytometries, Sortings, Molecular Biology, LcpIII, Protein Glycosylation, protein complex, malignant, Metastases, Apoptotic Bodies, Proteins, total expressed protein, enzyme activity, Flow Cytometries, Naevocarcinoma., Tumorigenesis, Cistrons, Cell, impact-a, Bioinformatics, Vesicle, Concept, O linked Glycosylation, Metastase, Melanomas, LC/MS/MS, beta 1-6 GlcNAc transferase V, native protein, natural protein, Malignant Melanoma, Protein, chemical analysis, proteomic analysis, Neoplasm, GlcNAcylations, IMPACT, imprinted and ancient gene protein homolog, Flow Microfluorometry, LCP3, Exovesicle, UDP-GLCNAC-alpha-mannoside beta(1, 4-N-acetylglucosaminyltransferase, polisacaridos, DMLCP3, l(2)28-28-12, body surface, UDP-N-acetylglucosamine-beta-D-mannoside beta-1, UDPGNAC-glycopeptide beta4-N-acetylglucosaminyl transferase III, Carcinogeneses, anon-35Fa, Cell Adhesions, N-acetylglucosaminyltransferase III, Age symptoms begin, Body, Protein Gene Products, DmelCG5861, Gene Proteins, Therapeutic, anon-35Fc, Sorting, liquid chromatography tandem mass spectrometry, Treatment, malignant melanoma, CG2043, assay, hereditary, RWDD5"],"name_synonyms":["Exovesicles, Apoptotic Body, Bodies, protein complex, Proteins, Apoptotic Bodies, Vesicles, Gene, proteins, protein, Extracellular Vesicle, protein-containing complex, cargo, Body, Protein Gene Products, Vesicle, Gene Proteins, protein polypeptide chains, native protein, natural protein, polypeptide chain, Extracellular, Protein, Apoptotic, Gene Products, Cell., metastatic melanoma, protein aggregate, metastatic malignant melanoma, Exovesicle"],"pubmed_title_synonyms":["Exovesicles, Apoptotic Body, Bodies, protein complex, Proteins, Apoptotic Bodies, Vesicles, Gene, proteins, protein, Extracellular Vesicle, protein-containing complex, cargo, Body, Protein Gene Products, Vesicle, Gene Proteins, protein polypeptide chains, native protein, natural protein, polypeptide chain, Extracellular, Protein, Apoptotic, Gene Products, Cell., metastatic melanoma, protein aggregate, metastatic malignant melanoma, Exovesicle"],"description_synonyms":["GNT-III protein, liquid chromatography tandem mass spectroscopy, Glc-N-Ac-transferase V, Materials, O-linked Glycosylations, determination, Metastasis, 6 N-acetylglucosaminyltransferase V, Galactosylation, DELTA AND GAMMA, Galactosylations, Glykan, Neoplasm Metastases, Extracellular Vesicle, protein, neoplasm metastasis, betaTub1, cargo, Molecular Biologies, cancer metastasis, polisacarido, rat, Techniques, protein polypeptide chains, B1t, Bio Informatics, 6)N-acetylglucosaminyltransferase, melanoma, GnT-III protein, Roles, Method, Concepts, 3, ramiform, Flow Microfluorimetry, protein aggregate, Cell Sortings, integumentum commune, imprinted and ancient gene protein, treatment, LCMSMS, kus, beta1tub, enzymes, CG9277, Glycans, Fluorescence Activated Cell Sorting, Biology, A330041C17Rik, N-Acetylglucosaminylation, T, beta-particle, Glycane, Age of onset, Oncogeneses, proteins, ramified, procedures, Bio-Informatic, Sialylation, Computational, alpha-1, genetic, 1t, Adhesions, e, e-, Methodological Studies, beta1Tub, Bio-Informatics, O-linked, gnt, Adhesion, Glycan, Role Concepts, disease management, Therapies, CG7776, Computational Molecular Biologies, N-Acetylglucosaminylations, Flow Cytofluorometry, Mgat5 protein, LC-MS2, Therapy, Cell Sorting, N-glycosyl-oligosaccharide-glycoprotein N-acetylglucosaminyltransferase V, BETA 56D, Fluorescence-Activated Cell Sorting, external covering of organism, statistics and numerical data, Biologies, mouse, e(Pc), familial, LC-MS/MS, beta-1, Glycosylations, organism surface, Malignant Melanomas, Procedure, L[[3]]CP3, Fucosylations, Computational Molecular, Glykane, Microfluorimetry, Melanoma, Role Concept, l(1)G0108, UDP-N-acetylglucosamine alpha-6-D-mannoside beta1, mannosyl (beta-1, Phosphoglycosylations, Playthings and Play, Apoptotic, Computational Molecular Biology, Role, Plaything, Genetic Materials, numerical data, FLOWERING TIME CONTROL PROTEIN FCA ALPHA, Genetic Material, beta-tub, Cytofluorometries, DmelCG7776, Beta <eudicots>, Bioinformatic, N linked Glycosylation, Toys, Sialylations, FCAALL.331, Methodological, beta56D, DmelCG9277, Methodological Study, Fluorescence-Activated Cell, Treatments, human, MGAT3 protein, beta1-tub, beta(-), DTB2, polysaccharides, GnT V enzyme, beta-Tub, 4-)-glycoprotein beta-1, Mgat3 protein, Oncogenesis, integumentary system, Material, N Acetylglucosaminylation, liquid chromatography-tandem mass spectroscopy, DmelLcp3, E(PC), i6, DL4180C, BG:DS02740.11, anon-48Ac, Biocatalyst, Cistron, inherited genetic, beta-Tub56D, E430016J11Rik, dermal system, Proteomes, betaTub, Specbeta, beta1-Tubulin, 4-N-acetylglucosaminyltransferase III, GH05739, Tumorigeneses, Plays, Exovesicles, GlcNAcylation, Procedures, Apoptotic Body, Bodies, Peptidomics, Biocatalysts, e(-), number, Vesicles, glycans, Gene, N-linked Glycosylations, electron, protein-containing complex, Malignant, Ximpact, LC-MS-MS, N-linked, Fluorescence-Activated, melanoma (disease), surface, Fluorescence-Activated Cell Sortings, Microfluorometry, O-linked Glycosylation, mannoside acetylglucosaminyltransferase 3, III, glycosylation, B-spec, N-acetylglucosaminyltransferase V, GH14582, polypeptide chain, Glycosylation, utilization, betaspec, 3(6)-mannosylglycoprotein beta-1, LC-MSMS, Studies, Gene Products, single organism cell adhesion, Technique, Polysaccharide, Cytofluorometry, Toy, DmelCG2043, study, tumor cell migration, betatub(56D), lcp3, Genetic, Playthings, Flow Cytofluorometries, Protein Glycosylations, cell adhesion molecule activity, Microfluorometries, Elektron, Naevocarcinoma, beta[[1]] tubulin, LCP-3, beta1t, metastasis, mannoside acetyl glucosaminyltransferase 3, anon-EST:fe1B3, Study, Enzyme, CP3, Puppets, SPEC8, Phosphoglycosylation, 6-N-acetylglucosaminyltransferase, Extracellular, Flow, Flow Microfluorometries, Cytometry, Play, CG5870, Fucosylation, constitutitional genetic, tumor metastasis, UDPGNAC-MAGtransferase III, N-linked Glycosylation, Puppet, Cytometries, Sortings, beta-tubulin56D, Spec-beta, Dmbeta1, Molecular Biology, use, LcpIII, Protein Glycosylation, protein complex, malignant, Metastases, Apoptotic Bodies, Proteins, total expressed protein, Tubulin, enzyme activity, Flow Cytometries, Naevocarcinoma., Tumorigenesis, i168, Cistrons, Cell, impact-a, Bioinformatics, Vesicle, Concept, O linked Glycosylation, Metastase, Melanomas, LC/MS/MS, beta 1-6 GlcNAc transferase V, native protein, natural protein, Malignant Melanoma, Protein, chemical analysis, proteomic analysis, Neoplasm, GlcNAcylations, betaSpec, IMPACT, imprinted and ancient gene protein homolog, techniques, Flow Microfluorometry, LCP3, b-Spec, Exovesicle, UDP-GLCNAC-alpha-mannoside beta(1, 4-N-acetylglucosaminyltransferase, DmelCG5870, polisacaridos, l(1)G0074, l(1)G0198, negatron, DMLCP3, beta[[1]]-tubulin, l(2)28-28-12, body surface, UDP-N-acetylglucosamine-beta-D-mannoside beta-1, UDPGNAC-glycopeptide beta4-N-acetylglucosaminyl transferase III, Carcinogeneses, b spectrin, anon-35Fa, beta1, Cell Adhesions, N-acetylglucosaminyltransferase III, Age symptoms begin, Body, beta, Tub, Protein Gene Products, DmelCG5861, Gene Proteins, Therapeutic, beta-spec, anon-35Fc, cardinality, BETA, Sorting, liquid chromatography tandem mass spectrometry, Treatment, malignant melanoma, CG2043, assay, hereditary, Spec, RWDD5, methodology"],"additional_accession":["PXD057657"]},"is_claimable":false,"name":"MGAT3 and MGAT5 overexpression alters the protein cargo of extracellular vesicles released by metastatic melanoma cells","description":"Extracellular vesicles (EVs) are potential non-invasive diagnostic, prognostic, and therapeutic tools. Additionally, they are important contributors to tumorigenesis. Glycosylation has been found to modulate the composition of the EV proteome, and increased amounts of beta 1,6-branched N-glycans, synthesized by N-acetylglucosaminyltransferase V (GnT-V), are most commonly observed in melanoma and associated with decreased cell adhesion and increased metastasis. The opposite effect is caused by the addition of bisecting GlcNAc by N-acetylglucosaminyltransferase III (GnT-III). To date, the impact of these enzymes on EV cargo in melanoma remains unexplored. Methods: Flow cytometry was used to study the surface glycosylation of genetic variants of WM266-4 melanoma cells with induced overexpression of GnT-III or GnT-V encoding genes (MGAT3 or MGAT5) and EVs released by these cells. LC-MS/MS proteomics was applied to analyze the effect of altered glycosylation on the proteome of released EVs, followed by thorough bioinformatic analysis. Results: Flow cytometry analysis showed dynamic changes in surface glycosylation of EVs derived from melanoma cells overexpressing MGAT3 or MGAT5. Induced MGAT3 or MGAT5 overexpression also resulted in significant changes in EVs proteome. Proteomic analysis identified the total number of 1770 microvesicular and 704 exosomal proteins that play diverse roles in melanoma progression, including those with established diagnostic/prognostic potential and those closely linked to melanoma onset. Conclusions: Proteomic profiling of EVs derived from MGAT3 and MGAT5 overexpressing cells revealed functional shifts in EV protein content driven by glycosylation modifications. The study presented a potential multidimensional use of melanoma-derived EVs for diagnostic and prognostic purposes. ","dates":{"publication":"Fri Nov 08 01:49:00 GMT 2024"},"accession":"MSV000096340","cross_references":{"pubmed":["40199132"]}}