Plant Proteostasis Methods and Protocols

This volume is a collection of detailed protocols describing state-of-art approaches that will facilitate the understanding of protein homeostasis in plant stress responses and development. Plant Proteostasis: Methods and Protocols is broken into four parts focusing on the study of ubiquitin-dependent post-translational modifications, protocols focused on Ubl post-translational modifications, protein homeostasis, and protocols for the in silico analysis. 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.

Authoritative and cutting-edge, Plant Proteostasis: Methods and Protocols aims to address next challenges in agriculture such as precision horticulture. 

Approaches to Determine Protein Ubiquitination Residue Types.- Immunoprecipitation of Cullin Ring Ligases (CRLs) in Arabidopsis Thaliana Seedlings.- Radioligand Binding Assays for Determining Dissociation Constants of Phytohormone Receptors.- Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.- Fluorescent Reporters for Ubiquitin-Dependent Proteolysis in Plants.- Generation of Artificial N-end Rule Substrate Proteins In Vivo and In Vitro.- Peptide Arrays for Binding Studies of E3 Ubiquitin Ligases.- SUMO Chain Formation by Plants Enzymes.- Kinetic Analysis of Plant SUMO Conjugation Machinery.- Expression, Purification, and Enzymatic Analysis Of Plant SUMO Proteases.- Quantitative Analysis of Subcellular Distribution of the SUMO Conjugation System by Confocal Microscopy Imaging.- Biochemical Analysis of Autophagy in Algae and Plants by Monitoring the Electrophoretic Mobility of ATG8.- Detection of Autophagy in Plants by Fluorescence Microscopy.- Protocols for Studying Protein Stability in a Arabidopsis Protoplast Transient Expression System.- Detection and Quantification of Protein Aggregates in Plants.- Determination of Protein Carbonylation and Proteasome Activity in Seeds.- Isobaric Tag for Relative and Absolute Quantitation (iTRAQ)-based Protein Profiling in Plants.-Use of a Phosphatidylinositol phosphate Affinity Chromatography (PIP - chromatography) for the Isolation of Proteins Involved in Protein Quality Control and Proteostasis Mechanisms in Plants.- In vivo Radiolabeling of Arabidopsis Leaves and Separation of Thylakoid Membrane Complexes by Blue Native PAGE.- Normalized Quantitative Western Blotting Based on Standardized Fluorescent Labeling.- Sequence Search and Comparative Genomic Analysis of SUMO-Activating Enzymes using CoGe.- Studying Evolutionary Dynamics of Gene Families Encoding SUMO-Activating Enzymes with SeaView and ProtTest.- Bioinformatics Tools for Exploring the SUMO Gene Network.