Abstract
Infection of cells and tissues by pathogenic microorganisms often involves severe reprogramming of host cell signaling. Typically, invasive microorganisms manipulate host cellular pathways seeking advantage for replication and survival within the host, or to evade the immune response. Understanding such subversion of the host cell by intracellular pathogens at a molecular level is the key to possible preventive and therapeutic interventions on infectious diseases. Reverse Protein Arrays (RPAs) have been exploited in other fields, especially in molecular oncology. However, this technology has not been applied yet to the study of infectious diseases. Coupling classic in vitro infection techniques used by cellular microbiologists to proteomic approaches such as RPA analysis should provide a wealth of information about how host cell pathways are manipulated by pathogens. The increasing availability of antibodies specific for phosphorylated epitopes in signaling proteins allows monitoring global changes in phosphorylation through the infection process by utilizing RPA analyses. In our lab, we have shown the potential of RPA technology in this field by devising a microarray consisting of lysates from cell cultures infected by Salmonella typhimurium. The protocols used are described here.
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Acknowledgments
We thank C. Molero, I. Rodríguez-Escudero, A. Alemán, R. Rotger, and other members of our lab for their help and support. M. Ehrat, G.M. Kresbach, and J. van Oostrum are acknowledged for their critical comments on the manuscript. This work was possible thanks to Grants BIO2007-67299 from Ministerio Educación y Ciencia and S-SAL-0246-2006 from Comunidad Autónoma de Madrid (Spain) to M. M.
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Cid, V.J., Kauffmann, E., Molina, M. (2011). Reverse Protein Arrays Applied to Host–Pathogen Interaction Studies. In: Wu, C. (eds) Protein Microarray for Disease Analysis. Methods in Molecular Biology, vol 723. Humana Press. https://doi.org/10.1007/978-1-61779-043-0_4
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DOI: https://doi.org/10.1007/978-1-61779-043-0_4
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