Discovery of Host–Viral Protein Complexes During Infection
Open accessAbstract
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.
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Affinity purification coupled to mass spectrometry allows a protein's interacting partners to be identified with little to no prior knowledge.
“Their integration with MS (AP-MS) (Figure 2) has allowed a protein’s interacting partners to be identified with little to no a priori knowledge (33).”
From The Impact of Mass Spectrometry–Based Proteomics on Fundamental Discoveries in Virology (2014) · cites this paper as [33]
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The Impact of Mass Spectrometry–Based Proteomics on Fundamental Discoveries in Virology
Greco, T. M., Diner, B. A., & Cristea, I. M. (2014) · Annual Review of Virology · doi:10.1146/annurev-virology-031413-085527 · Cited by 67 · see citing works
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- Authors
- Rowles, D. L., Terhune, S. S., & Cristea, I. M.
- Affiliations
- Princeton University (US)
- Medical College of Wisconsin (US)
- Corresponding author
- Terhune, Scott S.
- Cristea, Ileana M.
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2013
- Volume
- 1064
- Pages
- 43-70
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC4162421
- OpenAlex
- W32423614
- Cited by
- 15
- References
- 26
- Citations by year
- 2025: 2 · 2020: 2 · 2019: 3 · 2017: 1 · 2015: 4 · 2014: 3
- FWCI
- 20.45
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute on Drug Abuse (DP1DA026192)
- National Institute of Allergy and Infectious Diseases (R21AI102187, R01AI083281)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Health Sciences › Medicine › Epidemiology › Cytomegalovirus and herpesvirus research
- Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
- Keywords
- host-virus protein interaction, host-virus coevolution, viral protein function, immunoaffinity purification, mass spectrometry-based proteomics, DNA viruses, RNA viruses, human cytomegalovirus, herpes simplex virus type 1, pseudorabies virus, HIV-1, Sindbis virus, West Nile virus
- MeSH terms
- Animals, Chromatography, Affinity, Humans, Infections, Viral Proteins, Viruses, Protein Interaction Mapping, Proteomics, Multiprotein Complexes, Host-Pathogen Interactions
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