Ulteriori informazioni
Thisvolume is a collection of detailed protocols describing state-of-art approachesthat will facilitate the understanding of protein homeostasis in plant stressresponses and development. Plant Proteostasis: Methods and Protocols is broken into four parts focusing on thestudy of ubiquitin-dependent post-translational modifications, protocolsfocused on Ubl post-translational modifications, protein homeostasis, andprotocols for the in silico analysis. Written in the highly successful Methods in Molecular Biology seriesformat, chapters include introductions to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Plant Proteostasis: Methods andProtocols aims to address next challenges in agriculture such asprecision horticulture.
Sommario
Approaches to Determine Protein Ubiquitination ResidueTypes.- Immunoprecipitation of Cullin Ring Ligases(CRLs) in Arabidopsis Thaliana Seedlings.- Radioligand Binding Assays for Determining Dissociation Constants ofPhytohormone 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 ChainFormation by Plants Enzymes.- Kinetic Analysis of Plant SUMO Conjugation Machinery.- Expression, Purification, and EnzymaticAnalysis Of Plant SUMO Proteases.- Quantitative Analysis ofSubcellular Distribution of the SUMO Conjugation System by Confocal MicroscopyImaging.- Biochemical Analysis of Autophagy in Algae and Plants by Monitoringthe Electrophoretic Mobility of ATG8.- Detection of Autophagy inPlants 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 ProteinCarbonylation and Proteasome Activity in Seeds.- Isobaric Tag for Relativeand Absolute Quantitation (iTRAQ)-based Protein Profiling in Plants.-Use of a Phosphatidylinositol phosphate AffinityChromatography (PIP - chromatography) for the Isolation of Proteins Involved inProtein Quality Control and Proteostasis Mechanisms in Plants.- In vivo Radiolabeling of Arabidopsis Leaves and Separation of ThylakoidMembrane Complexes by Blue Native PAGE.- Normalized Quantitative Western BlottingBased on Standardized Fluorescent Labeling.- Sequence Search andComparative Genomic Analysis of SUMO-Activating Enzymes using CoGe.- Studying Evolutionary Dynamicsof Gene Families Encoding SUMO-Activating Enzymes with SeaView and ProtTest.- Bioinformatics Tools for Exploring the SUMO Gene Network.
Riassunto
Plant Proteostasis: Methods and Protocols is broken into four parts focusing on thestudy of ubiquitin-dependent post-translational modifications, protocolsfocused on Ubl post-translational modifications, protein homeostasis, andprotocols for the in silico analysis.
