Author: Munday, Diane C.; Emmott, Edward; Surtees, Rebecca; Lardeau, Charles-Hugues; Wu, Weining; Duprex, W. Paul; Dove, Brian K.; Barr, John N.; Hiscox, Julian A.
Title: Quantitative Proteomic Analysis of A549 Cells Infected with Human Respiratory Syncytial Virus Document date: 2010_7_20
ID: 2zhaknbi_5
Snippet: To take an unbiased, global approach to investigating changes in the host cell proteome in response to HRSV infection, stable isotope labeling with amino acids in cell culture (SILAC) was used in conjunction with LC-MS/MS. Through this method, cellular and possibly viral proteins were identified and quantified. To reduce sample complexity and to study the interaction of HRSV with different regions of the cell, nuclear and cytoplasmic fractions we.....
Document: To take an unbiased, global approach to investigating changes in the host cell proteome in response to HRSV infection, stable isotope labeling with amino acids in cell culture (SILAC) was used in conjunction with LC-MS/MS. Through this method, cellular and possibly viral proteins were identified and quantified. To reduce sample complexity and to study the interaction of HRSV with different regions of the cell, nuclear and cytoplasmic fractions were purified and analyzed. A549 cells, a human lung carcinoma cell line that has properties similar to those of alveolar cells (which are permissive for HRSV), were used in this study. Because of its respiratory origin, this cell line has been used extensively in the characterization of HRSV infection (18, 21, 24, 28 -30) and in the proteomic analysis of cellular and infectious respiratory diseases (26, (31) (32) (33) . The quantitative proteomic analysis was validated using alternative techniques such as Western blot, confocal microscopy, and functional assays where appropriate. The data generated in this study indicated that there were cellular proteome alterations in HRSV-infected cells when compared with mock-infected cells and that many alterations were specific to defined protein subsets, pathways, and organelles. These included mitochondrial proteins, cell cycle regulatory proteins, ND10s, components of the nuclear pore complex, and nucleocytoplasmic trafficking proteins.
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