title: Characterizing and measuring endocytosis of lipid-binding effectors in mammalian cells.
abstract: Pathogen-host interactions are mediated in part by secreted microbial proteins capable of exploiting host cells for their survival. Several of these manipulations involve, but are not limited to, suppression of defense responses, alterations in host vesicular trafficking, and manipulation of gene expression. The delivery of such molecules from microbe to host has been of intense interest in several microbe-host systems. Several well-studied bacterial effectors are delivered directly into host cells through a needle injection apparatus. Conversely, there have been several examples of secreted effectors and protein toxins from bacteria and eukaryotic microbes, such as fungi and oomycetes, being internalized into host cells by receptor-mediated endocytosis. In the following chapter, we discuss various techniques utilized to measure these endocytosed lipid-binding effectors that can be delivered in the absence of the pathogen.
authors.affiliation: Virginia Bioinformatics Institute, Virginia Tech, Blacksburg, Virginia, USA; Department of Biochemistry, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Helen R
authors.initials: HR
authors.lastname: Clark
authors.affiliation: Virginia Bioinformatics Institute, Virginia Tech, Blacksburg, Virginia, USA; Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Tristan A
authors.initials: TA
authors.lastname: Hayes
authors.affiliation: Virginia Bioinformatics Institute, Virginia Tech, Blacksburg, Virginia, USA. Electronic address: sdkale@vt.edu.
authors.forename: Shiv D
authors.initials: SD
authors.lastname: Kale
mesh_terms: D001665:Binding Sites; D002460:Cell Line; D002846:Chromatography, Affinity; D004705:Endocytosis; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D054884:Host-Pathogen Interactions; D006801:Humans; D018613:Microscopy, Confocal; D010838:Phytophthora; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection
chemical_list: D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: Cell entry; Confocal microscopy; Effectors; Flow cytometry; Phospholipids