Identifying the Membrane Proteome of HIV-1 Latently Infected Cells

Profiling integral plasma membrane proteins is of particular importance for the identification of new biomarkers for diagnosis and for drug development. We report in this study the identification of surface markers by performing comparative proteomics of established human immunodeficiency virus-1 (H...

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Published in:The Journal of biological chemistry 2007-03, Vol.282 (11), p.8207-8218
Main Authors: Berro, Reem, de la Fuente, Cynthia, Klase, Zachary, Kehn, Kylene, Parvin, Lida, Pumfery, Anne, Agbottah, Emmanuel, Vertes, Akos, Nekhai, Sergei, Kashanchi, Fatah
Format: Article
Language:English
Subjects:
Agammaglobulinaemia Tyrosine Kinase
Apoptosis
Blotting, Western
Cell Line
Cell Membrane - metabolism
Cell Membrane - virology
Cell Survival
HIV-1 - metabolism
Human immunodeficiency virus 1
Humans
Leukocytes, Mononuclear - metabolism
Leukocytes, Mononuclear - virology
Models, Biological
Phosphatidylinositol 3-Kinases - metabolism
Protein-Tyrosine Kinases - metabolism
Proteomics - methods
Viral Proteins - chemistry
X-Linked Inhibitor of Apoptosis Protein - metabolism
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Summary:Profiling integral plasma membrane proteins is of particular importance for the identification of new biomarkers for diagnosis and for drug development. We report in this study the identification of surface markers by performing comparative proteomics of established human immunodeficiency virus-1 (HIV-1) latent cell models and parental cell lines. To this end we isolated integral membrane proteins using a biotin-directed affinity purification method. Isolated proteins were separated by two-dimensional gel electrophoresis and identified by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) after in gel digestion. Seventeen different proteins were found to vary on the surface of T-cells due to HIV-1 infection. Of these proteins, 47% were integral membrane proteins, and 18% were membrane-associated. Through the use of complementary techniques such as Western blotting and fluorescent staining, we confirmed the differential expression of some of the proteins identified by MALDI-TOF including Bruton's tyrosine kinase and X-linked inhibitor of apoptosis. Finally, using phosphatidylinositol 3-kinase inhibitors and flavopiridol to inhibit Bruton's tyrosine kinase localization at the membrane and X-linked inhibitor of apoptosis protein expression, respectively, we showed that HIV-1 latently infected cells are more sensitive to these drugs than uninfected cells. This suggests that HIV-1 latently infected cells may be targeted with drugs that alter several pathways that are essential for the establishment and maintenance of latency.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M606324200