title: Proteomics characterization of exosome cargo.
abstract: Characterization of exosomal cargo is of significant interest because this cargo can provide clues to exosome biogenesis, targeting, and cellular effects and may be a source of biomarkers for disease diagnosis, prognosis and response to treatment. With recent improvements in proteomics technologies, both qualitative and quantitative characterization of exosomal proteins is possible. Here we provide a brief review of exosome proteomics studies and provide detailed protocols for global qualitative, global quantitative, and targeted quantitative analysis of exosomal proteins. In addition, we provide an example application of a standard global quantitative analysis followed by validation via a targeted quantitative analysis of urine exosome samples from human patients. Advantages and limitations of each method are discussed as well as future directions for exosome proteomics analysis.
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, United States. Electronic address: k.schey@vanderbilt.edu.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Schey
authors.affiliation: Department of Medicine, Vanderbilt University School of Medicine, United States; Department of Pharmacology, Vanderbilt University School of Medicine, United States.
authors.forename: J Matthew
authors.initials: JM
authors.lastname: Luther
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, United States.
authors.forename: Kristie L
authors.initials: KL
authors.lastname: Rose
mesh_terms: D000595:Amino Acid Sequence; D017302:Annexins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015703:Antigens, CD / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015706:Bibliometrics; D001692:Biological Transport; D015415:Biomarkers / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D055354:Exosomes / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000099177:Tsg101 Protein
chemical_list: D017302:Annexins; D015703:Antigens, CD; D015415:Biomarkers; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D020543:Proteome; D014157:Transcription Factors; D000099177:Tsg101 Protein
keywords: Mass spectrometry; Post-translational modifications; Proteomics
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