title: Proteomic Characterization of Exosomes from HIV-1-Infected Cells.
abstract: Proteomics has increasingly become an invaluable tool to characterize proteomes from various subcellular compartments. Here, we describe a quantitative proteomics method using the technique of Stable Isotope Labeling by Amino acids in Cell culture (SILAC) to analyze the effects of HIV infection on host exosomal proteomes. The procedure, described below, involves differential isotope labeling of cells, exosome purification, mass spectrometric quantification, and various bioinformatic analyses/verifications. Although this chapter focuses on analyzing the effects of HIV-1 infection on the exosomal proteome, the protocol can easily be adapted to other subcellular compartments under different stress conditions.
authors.affiliation: Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, The Warren Alpert Medical School of Brown University, 55 Claverick Street, Room 430, Providence, RI, 02903, USA. ming.li@brown.edu.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, The Warren Alpert Medical School of Brown University, 55 Claverick Street, Lab 414, Providence, RI, 02903, USA. BRamratnam@lifespan.org.
authors.forename: Bharat
authors.initials: B
authors.lastname: Ramratnam
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D055354:Exosomes / Q000378:metabolism* / Q000821:virology; D015658:HIV Infections / Q000378:metabolism* / Q000821:virology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
chemical_list: D011506:Proteins
keywords: Exosome; HIV-1; Mass spectrometry; Proteomics; SILAC