title: Chemoenzymatic site-specific labeling of influenza glycoproteins as a tool to observe virus budding in real time.
abstract: The influenza virus uses the hemagglutinin (HA) and neuraminidase (NA) glycoproteins to interact with and infect host cells. While biochemical and microscopic methods allow examination of the early steps in flu infection, the genesis of progeny virions has been more difficult to follow, mainly because of difficulties inherent in fluorescent labeling of flu proteins in a manner compatible with live cell imaging. We here apply sortagging as a chemoenzymatic approach to label genetically modified but infectious flu and track the flu glycoproteins during the course of infection. This method cleanly distinguishes influenza glycoproteins from host glycoproteins and so can be used to assess the behavior of HA or NA biochemically and to observe the flu glycoproteins directly by live cell imaging.
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Maximilian Wei-Lin
authors.initials: MW
authors.lastname: Popp
authors.forename: Roos A
authors.initials: RA
authors.lastname: Karssemeijer
authors.forename: Hidde L
authors.initials: HL
authors.lastname: Ploegh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D004285:Dogs; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D013194:Staining and Labeling / Q000379:methods; D014771:Virion / Q000201:enzymology; D019065:Virus Assembly / Q000502:physiology*; D057074:Virus Release / Q000502:physiology*
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D009439:Neuraminidase