title: Affinity Purification-Mass Spectroscopy Methods for Identifying Epstein-Barr Virus-Host Interactions.
abstract: Considerable insight into the function and mechanism of action of viral proteins has come from identifying the cellular proteins with which they interact. In recent years, mass spectrometry-based methods have emerged as the method of choice for protein interaction discovery due to their comprehensive and unbiased nature. Methods involving single affinity purifications of epitope-tagged viral proteins (AP-MS) and tandem affinity purifications of viral proteins with two purification tags (TAP tagging) have both been used to identify novel host interactions with EBV proteins. However, to date these methods have only been applied to a small number of EBV proteins. Here we provide detailed methods of AP-MS and TAP tagging approaches that can be applied to any EBV protein in order to discover its host interactions.
authors.affiliation: Department of Molecular Genetics, University of Toronto, 1 Kings College Circle, Toronto, ON, Canada, M5S 1A8.
authors.forename: Anna A
authors.initials: AA
authors.lastname: Georges
authors.affiliation: Department of Molecular Genetics, University of Toronto, 1 Kings College Circle, Toronto, ON, Canada, M5S 1A8. lori.frappier@utoronto.ca.
authors.forename: Lori
authors.initials: L
authors.lastname: Frappier
mesh_terms: D002846:Chromatography, Affinity; D020031:Epstein-Barr Virus Infections / Q000378:metabolism* / Q000821:virology*; D004854:Herpesvirus 4, Human / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D059625:Liquid-Liquid Extraction; D013058:Mass Spectrometry; D011485:Protein Binding; D020543:Proteome; D040901:Proteomics / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D020543:Proteome; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins
keywords: Affinity purification; Epstein–Barr virus; FLAG tag; Mass spectrometry; Protein interactions; Proteomics; SPA tag; TAP tag