Medical College of Wisconsin
CTSICores SearchResearch InformaticsREDCap

Discovery of host-viral protein complexes during infection. Methods Mol Biol 2013;1064:43-70

Date

09/03/2013

Pubmed ID

23996249

Pubmed Central ID

PMC4162421

DOI

10.1007/978-1-62703-601-6_4

Scopus ID

2-s2.0-84884176527 (requires institutional sign-in at Scopus site)   13 Citations

Abstract

Viruses have co-evolved with their hosts, developing effective approaches for hijacking and manipulating host cellular processes. Therefore, for their efficient replication and spread, viruses depend on dynamic and temporally regulated interactions with host proteins. The rapid identification of host proteins targeted by viral proteins during infection provides significant insights into mechanisms of viral protein function. The resulting discoveries often lead to unique and innovative hypotheses on viral protein function. Here, we describe a robust method for identifying virus-host protein interactions and protein complexes, which we have successfully utilized to characterize spatial-temporal protein interactions during infections with either DNA or RNA viruses, including human cytomegalovirus (HCMV), herpes simplex virus type 1 (HSV-1), pseudorabies virus (PRV), human immunodeficiency virus (HIV-1), Sindbis, and West Nile virus (WNV). This approach involves cryogenic cell lysis, rapid immunoaffinity purification targeting a virus or host protein, followed by identification of associated proteins using mass spectrometry. Like most proteomic approaches, this methodology has evolved over the past few years and continues to evolve. We are presenting here the updated approaches for each step, and discuss alternative strategies allowing for the protocol to be optimized for different biological systems.

Author List

Rowles DL, Terhune SS, Cristea IM

Author

Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Chromatography, Affinity
Host-Pathogen Interactions
Humans
Multiprotein Complexes
Protein Interaction Mapping
Proteomics
Viral Proteins
Viruses