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This detailed volume introduces different methods to assess the structure and folding of bacterial amyloids and analyze their functions, either in vitro or in vivo. Despite their initial association with neurodegenerative diseases, there is now increasing evidence of alternative roles for amyloids, with beneficial aspects for cells. In particular, prokaryotes use amyloids as functional assemblies, where they are key players in the cell's physiology. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Bacterial Amyloids: Methods and Protocols is an ideal guide for experts and novices alike to further explore these crucial protein assemblies.
Sommario
Study of Amyloid Fibers Using Atomic Force Microscopy.- Cryo-Electron Microscopy to Analyze the Structure of Bacterial Amyloids In Vitro.- Monitoring Molecular Assembly of Biofilms Using Quartz Crystal Microbalance with Dissipation (QCM-D).- Measuring the Size and Spontaneous Fluctuations of Amyloid Aggregates with Fluorescence Correlation Spectroscopy.- Determining the Stoichiometry of Amyloid Oligomers by Single Molecule Photobleaching.- Structural Information on Bacterial Amyloid and Amyloid-DNA Complex Obtained by Small-Angle Neutron/X-Ray Scattering.- Fiber Diffraction and Small-Angle Scattering for Structural Investigation of Bacterial Amyloids.- Supported Lipid Bilayers (SLBs) to Study Amyloid-Lipid Membrane Interactions with Atomic Force Microscopy.- Characterization of Bacterial Amyloids by Nano-Infrared Spectroscopy.- High Throughput Screening of Heterologous Functional Amyloids Using Escherichia coli.- Evaluation of Amyloid Inhibitor Efficiency to Block Bacterial Survival.- Anti-Amyloid Drug Screening Methods Using Bacterial Inclusion Bodies.- Determination of Effects and Mechanisms of Action of Bacterial Amyloids on Antibiotic Resistance.- Design and Testing of Synthetic Catalytic Amyloids Based on the Active Site of Enzymes.- Analysis of Bacterial Amyloid Interaction with Lipidic Membrane by Orientated Circular Dichroism and Infrared Spectroscopies.- Direct, Rapid, and Simple Evaluation of the Expression and Conformation of Beta-Amyloid in Bacterial Cells by FTIR Spectroscopy.- Assessment of Intracellular Amyloid Formation in Fixed and Live Bacteria Using Fluorescence Microscopy.- Identification of Aggregation-Prone and Gatekeeper Residues in Bacterial Amyloids Using Site-Directed Mutagenesis and Flow Cytometry.- Probing DNA-Amyloid Interaction and Gel Formation by Active Magnetic Wire Microrheology.- Probing Amyloids-DNA Interaction with Nanofluidics.- Evaluation of the Role of Bacterial Amyloid on Nucleoid Structure Using Cryo Soft X-Ray Tomography.
