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Proteomic Characterization of Exosomes from HIV-1-Infected Cells

  • Protocol
HIV Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1354))

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.

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Acknowledgements

This work was supported by an ARRA supplement to the Lifespan/Tufts/Brown CFAR, P30AI042853-13S1, NIH P20GM103421, P01AA019072, R01HD072693 to BR. This work was also supported by Lifespan Pilot Research Fund (#701-5857), Rhode Island Foundation Medical Research Grant (#20133969), and NIH COBRE URI/RIH Pilot Research Grant (P20GM104317) to ML.

Author information

Authors and Affiliations

  1. 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 Ph.D.

  2. 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

    Bharat Ramratnam M.D.

Authors
  1. Ming Li Ph.D.
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  2. Bharat Ramratnam M.D.
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Corresponding authors

Correspondence to Ming Li Ph.D. or Bharat Ramratnam M.D. .

Editor information

Editors and Affiliations

  1. Department of Microbiology & Immunology, Albert Einstein College of Medicine, BRONX, New York, USA

    Vinayaka R. Prasad

  2. Department of Genetics, Albert Einstein College of Medicine, BRONX, New York, USA

    Ganjam V. Kalpana

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Li, M., Ramratnam, B. (2016). Proteomic Characterization of Exosomes from HIV-1-Infected Cells. In: Prasad, V., Kalpana, G. (eds) HIV Protocols. Methods in Molecular Biology, vol 1354. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3046-3_21

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  • DOI: https://doi.org/10.1007/978-1-4939-3046-3_21

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3045-6

  • Online ISBN: 978-1-4939-3046-3

  • eBook Packages: Springer Protocols

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