Quantitative Proteomics Using Stable Isotope Labeling with Amino Acids in Cell Culture Reveals Protein and Pathway Regulation in Porcine Circovirus Type 2 Infected PK-15 Cells

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• 作者：Huiying Fan ; Yu Ye ; Yongwen Luo ; Tiezhu Tong ; Guangrong Yan ; Ming Liao
• 刊名：Journal of Proteome Research
• 出版年：2012
• 出版时间：February 3, 2012
• 年：2012
• 卷：11
• 期：2
• 页码：995-1008
• 全文大小：574K
• 年卷期：v.11,no.2(February 3, 2012)
• ISSN：1535-3907

文摘

The infection of host cells by porcine circovirus type 2 (PCV2) leads to extensive modulation of the gene expression levels of target cells. To uncover the pathogenesis and virus-host interactions of PCV2, a quantitative proteomic study using the stable isotope labeling with amino acids in cell culture (SILAC), coupled with mass spectrometry, was performed on PCV2-infected PK-15 cells. The SILAC-based approach identified 1341 proteins, 163 of which showed significant change in level at 72 h after infection (79 up-regulated and 84 down-regulated). The modulated proteins included a number of proteins involved in substrate transport, cytoskeletal changes, and the stress response. Changes in the expression levels of selected proteins were verified by Western blot analysis. Ingenuity Pathway Analysis was used to reveal protein and interactive pathway regulation in response to PCV2 infection. Functional network and pathway analyses could provide insights into the complexity and dynamics of virus鈥揾ost cell interactions and may accelerate our understanding of the mechanisms of PCV2 infection.

Keywords:

porcine circovirus type 2; quantitative proteomics; stable isotope labeling with amino acids in cell culture (SILAC); pathway analysis