Immunoprecipitation Followed by Mass Spectrometry: An Approach for Identifying Host-Viral Protein-Protein Interactions

Abstract

As obligate intracellular parasites, viruses rely on the efficient manipulation of the cell they invade in order to multiply and spread. Protein-protein interactions between viral proteins (or their complexes) and cellular proteins are at the interface between virus and host and hence crucial for the outcome of the infection. Multiple techniques can be used to study protein-protein interactions in vivo in the context of the infected cell; among them, immunoprecipitation followed by mass spectrometry (IP-MS) has proven an efficient approach for the unbiased identification of protein complexes containing a viral protein of interest. In this chapter, we discuss how to employ IP-MS to define the interactome of plant virus proteins by using transient expression in the experimental host Nicotiana benthamiana, using the geminivirus tomato yellow leaf curl virus (TYLCV) as an example.

Keywords: IP-MS; Immunoprecipitation; Mass spectrometry; Nicotiana benthamiana; Protein complexes; Protein–protein interactions.