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Journal of Virology, October 2008, p. 9689-9699, Vol. 82, No. 19
0022-538X/08/$08.00+0     doi:10.1128/JVI.00995-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Quantitative Whole-Cell Proteome Analysis of Pseudorabies Virus-Infected Cells{triangledown}

Martin Skiba, Thomas C. Mettenleiter, and Axel Karger*

Institute of Molecular Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany

Received 13 May 2008/ Accepted 16 July 2008

A quantitative proteome study using the stable isotope labeling with amino acids in cell culture technique was performed on bovine kidney cells after infection with the alphaherpesvirus pseudorabies virus (PrV), the etiological agent of Aujeszky's disease. To enhance yields of proteins to be identified, raw extracts were fractionated by affinity solid-phase extraction with a combination of a cibacron blue F3G-A and a heparin matrix and with a phosphoprotein-specific matrix. After two-dimensional gel electrophoresis in different pH ranges between pH 3 and pH 10, 2,600 proteins representing 565 genes were identified by mass spectrometry and screened for virus-induced changes in relative protein levels. Four hours after infection, significant quantitative variations were found for constituents of the nuclear lamina, representatives of the heterogeneous nuclear ribonucleoproteins, proteins involved in membrane trafficking and intracellular transport, a ribosomal protein, and heat shock protein 27. Several proteins were present in multiple charge variants that were differentially affected by infection with PrV. As a common pattern for all these proteins, a mass shift in favor of the more acidic isoforms was observed, suggesting the involvement of viral or cellular kinases.

* Corresponding author. Mailing address: Institute of Molecular Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany. Phone: 49-38351-7251. Fax: 49-38351-7175. E-mail: axel.karger{at}fli.bund.de

{triangledown} Published ahead of print on 23 July 2008.

Journal of Virology, October 2008, p. 9689-9699, Vol. 82, No. 19
0022-538X/08/$08.00+0     doi:10.1128/JVI.00995-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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