title: Guanylate binding proteins directly attack Toxoplasma gondii via supramolecular complexes.
abstract: GBPs are essential for immunity against intracellular pathogens, especially for Toxoplasma gondii control. Here, the molecular interactions of murine GBPs (mGBP1/2/3/5/6), homo- and hetero-multimerization properties of mGBP2 and its function in parasite killing were investigated by mutational, Multiparameter Fluorescence Image Spectroscopy, and live cell microscopy methodologies. Control of T. gondii replication by mGBP2 requires GTP hydrolysis and isoprenylation thus, enabling reversible oligomerization in vesicle-like structures. mGBP2 undergoes structural transitions between monomeric, dimeric and oligomeric states visualized by quantitative FRET analysis. mGBPs reside in at least two discrete subcellular reservoirs and attack the parasitophorous vacuole membrane (PVM) as orchestrated, supramolecular complexes forming large, densely packed multimers comprising up to several thousand monomers. This dramatic mGBP enrichment results in the loss of PVM integrity, followed by a direct assault of mGBP2 upon the plasma membrane of the parasite. These discoveries provide vital dynamic and molecular perceptions into cell-autonomous immunity.
authors.affiliation: Institute of Medical Microbiology and Hospital Hygiene, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kravets
authors.affiliation: Institute of Medical Microbiology and Hospital Hygiene, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Degrandi
authors.affiliation: Institute for Molecular Physical Chemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Qijun
authors.initials: Q
authors.lastname: Ma
authors.affiliation: Institute for Molecular Physical Chemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Thomas-Otavio
authors.initials: TO
authors.lastname: Peulen
authors.affiliation: Institute of Medical Microbiology and Hospital Hygiene, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Klümpers
authors.affiliation: Institute for Molecular Physical Chemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Suren
authors.initials: S
authors.lastname: Felekyan
authors.affiliation: Institute for Molecular Physical Chemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Kühnemuth
authors.affiliation: Center of Advanced Imaging, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Weidtkamp-Peters
authors.affiliation: Institute for Molecular Physical Chemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Claus Am
authors.identifier: http://orcid.org/0000-0002-5171-149X
authors.initials: CA
authors.lastname: Seidel
authors.affiliation: Institute of Medical Microbiology and Hospital Hygiene, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Pfeffer
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D031541:Fluorescence Resonance Energy Transfer; D019204:GTP-Binding Proteins / Q000378:metabolism*; D051379:Mice; D008853:Microscopy; D061848:Optical Imaging; D055503:Protein Multimerization; D013052:Spectrometry, X-Ray Emission; D014122:Toxoplasma / Q000187:drug effects / Q000276:immunology* / Q000502:physiology*; D014617:Vacuoles / Q000469:parasitology
chemical_list: D019204:GTP-Binding Proteins; C452200:Gbp2 protein, mouse
keywords: Förster resonance energy transfer; biophysics; cell-autonomous immunity; guanylate-binding proteins; host-pathogen interaction; infectious disease; microbiology; mouse; multiparameter fluorescence image spectroscopy; structural biology
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