Bacterial Protein Secretion Systems

Identification of Protein Secretion Systems in Bacterial Genomes using MacSyFinder.- Protein Sorting Prediction .- Cell Fractionation.- Defining Lipoprotein Localisation by Fluorescence Microscopy.- Components Sub-cellular Localization: Identification of Lipoproteins using Globomycin and Radioactive Palmitate.- Defining Membrane Protein Localization by Isopycnic Density Gradients.- Components Sub-cellular Localisation: Cell Surface Exposure.- Probing Inner Membrane Protein Topology by Proteolysis.- Mapping of Membrane Protein Topology by Substituted Cysteine Accessibility Method (SCAMTM).- Defining Membrane Protein Topology using pho-lac Reporter Fusions.- In vivo and in vitro Protein-peptidoglycan Interactions.- Measure of Peptidoglycan Hydrolase Activity.- Protein-protein Interaction: Bacterial Two-Hybrid.- Protein-protein Interactions: Yeast two-hybrid.- Protein-protein Interactions: Cytology Two-hybrid.- Fusion Reporter Approaches to Monitor Transmembrane Helix Interactions in Bacterial Membranes.- Protein-protein Interactions: Co-immunoprecipitation.- Protein-protein Interaction: Tandem Affinity Purification in Bacteria.- Site-directed and Time-resolved Photocrosslinking in Cells Metabolically Labeled With Radioisotopes.- Protein-protein Interactions: Pull-down Assays.- Protein-protein interactions: Surface Plasmon Resonance.- Assessing Energy-dependent Protein Conformational Changes in the TonB System.- Defining Assembly Pathways by Fluorescence Microscopy.- Large Complexes: Cloning Strategy, Production, and Purification.- Shearing and Enrichment of Extracellular Type IV Pili.- Blue Native PAGE Analysis of Bacterial Secretion Complexes.- In situ Imaging of Bacterial Secretion Systems by Electron Cryo-tomography.- Structural Analysis of Protein Complexes by Cryo Electron Microscopy.- Bacterial Filamentous Appendages Investigated by Solid-state NMR Spectroscopy.- Identification of Effectors:  Precipitation of Supernatant Material.- Energy Requirements for Protein Secretion via the Flagellar type-III Secretion System.- Screening for Secretion of the Type VI Secretion Protein Hcp by Enzyme-Linked Immunosorbent Assay and Colony Blot.- Effector Translocation: Cya Reporter Assay.- Monitoring Effector Translocation with the TEM-1 Beta-lactamase Reporter System.- Effector Translocation Assay: Differential Solubilization.- Quantitative Determination of Anti-bacterial Activity during Bacterial Co-culture.