Plant Endosomes: Methods and Protocols

ISBN-13: 9781493914197 / Angielski / Twarda / 2014 / 286 str.

Marisa S. Otegui

Opis

Plant Endosomes: Methods and Protocols explores a collection of protocols and techniques to analyze in vivo trafficking of endocytic/endosomal cargo, including lipids, fluids, proteins and ligands, ultrastructural features of endosomes by high-pressure freezing/freeze-substitution and electron tomography, as well as protein-protein interactions in the endosomal and endomembrane system. The volume continues with coverage of the sorting defects in the transport of vacuolar storage proteins, function conservation of plant endosomal proteins, endosomal trafficking during plant responses to pathogens, protein composition of endosomes and endocytic vesicles, ubiquitination of endosomal cargo proteins and the identification of novel endosomal components by chemical genomics and proteomics. Written in the highly successful Methods in Molecular Biology series format, 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.Detailed and practical, Plant Endosomes: Methods and Protocols gathers contributions from many leading and emerging plant membrane trafficking researchers in order to promote and facilitate novel studies and ideas in this vital field.

Spis treści

The Use of FM Dyes to Analyze Plant Endocytosis.- Sterol Dynamics During Endocytic Trafficking in Arabidopsis.- Live Microscopy Analysis of Endosomes and Vesicles in Tip-Growing Root Hairs.- Analysis of Fluid Phase Endocytosis in (Intact) Plant Cells.- Immunogold Labeling and Electron Tomography of Plant Endosomes.- Investigating Protein-Protein Interactions in the Plant Endomembrane System Using Multiphoton-Induced FRET-FLIM.- Analysis of Rab GTPase-Effector Interactions by Bimolecular Fluorescence Complementation.- In Vivo Imaging of Brassinosteroid Endocytosis in Arabidopsis.- Analysis of Prevacuolar Compartment-Mediated Vacuolar Proteins Transport.- Evaluation of Defective Endosomal Trafficking to the Vacuole by Monitoring Seed Storage Proteins in Arabidopsis thaliana.- Trans-Species Complementation Analysis to Study Function Conservation of Plant Endosomal Sorting Complex Required for Transport (ESCRT) Proteins.- A Re-Elicitation Assay to Correlate flg22-Signaling Competency with Ligand-Induced Endocytic Degradation of the FLS2 Receptor.- Preparation of Enriched Plant Clathrin-Coated Vesicles by Differential and Density Gradient Centrifugation.- Proteomics of Endosomal Compartments from Plants Case Study: Isolation of trans-Golgi Network Vesicles.- Analysis of Global Ubiquitylation and Ubiquitin-Binding Domains Involved in Endosomal Trafficking.- Analysis of Endocytosis and Ubiquitination of the BOR1 Transporter.- Ubiquitination of Plant Immune Receptors.- Ubiquitylation-Mediated Control of Polar Auxin Transport: Analysis of Arabidopsis PIN2 Auxin Transport Protein.- Chemical Genomics Screening for Biomodulators of Endomembrane System Trafficking.- Integrative Chemical Proteomics and Cell Biology Methods to Study Endocytosis and Vesicular Trafficking in Arabidopsis.